نتائج البحث

This Was a Multinational Study Comparing the Efficacy and Safety of Two Medicines , Solifenacin Succinate and Mirabegron Taken Together, or Separately, or a Mock Treatment (Placebo) in Subjects With Symptoms of Overactive Bladder (SYNERGY)

• Source: A Randomized, Double-Blind, Parallel-Group, Placebo- and Active-Controlled, Multi-center Study to Evaluate the Efficacy, Safety and Tolerability of Combinations of Solifenacin Succinate and Mirabegron

تفاصيل العنوان

Movie 7: Live imaging of nuclei under ambient temperature (mock treatment)

• Authors : Dumur, Tao (Gregor Mendel Institute)

تفاصيل العنوان

Figure 3. EPF1 signaling downregulates both MUTE promoter activity and MUTE protein accumulation. ; (A) Representative confocal images of abaxial cotyledon epidermis from 4-day-old estradiol-inducible EPF1 seedlings carrying MUTEpro::nucYFP and MUTEpro::MUTE-GFP four days after mock-treated or estradiol-treated for induced EPF1 overexpression (iEPF1). F1 seedlings were used for the analysis. Scale bars, 25 µm. Experiments were repeated three times. Three seedlings were analyzed each time. (B) Representative confocal images of abaxial cotyledon epidermis from 4-day-old estradiol-inducible EPF1 seedlings carrying SCRMpro::nucGFP and SCRMpro::GFP-SCRM four days after mock-treated or estradiol-treated for induced EPF1 overexpression (iEPF1). F1 seedlings were used for the analysis. Scale bars, 25 µm. Experiments were repeated three times. Three seedlings were analyzed each time. (C) Representative Z-stack confocal image projection showing the MUTE-GFP levels after iEPF1 induction used for the quantitative analysis. Meristemoids accumulating MUTE-GFP are indicated by arrowheads. F1 seedlings were used for the analysis. Scale bars, 10 µm. Experiments were repeated three times. Three seedlings were analyzed each time. (D) Quantitative analysis of MUTE-GFP levels after iEPF1 induction at the time indicated on 3-day-old seedlings. Each dot represents the total intensity value of GFP signals from each nucleus expressing MUTE-GFP. To fully cover the entire nuclei expressing MUTE-GFP, serial Z-stack projection images were used for quantitative analysis (see Materials and methods). Mean value at each time point is connected by the line to visualize the average intensity change over time. Purple, initial values at time point 0; Cyan, mock treatment; Magenta, iEPF1 induction. Experiments were repeated three times; n = 6 for time point 0; n = 3 per subsequent time point. Total of 495 nuclei were analyzed. (E) Quantitative analysis of SCRMpro::nucGFP levels after iEPF1 induction at the time indicated on 3-day-old seedlings. Each dot represents the total intensity value of GFP signals from each nucleus expressing SCRMpro::nucGFP. To fully cover the entire nuclei expressing GFP, serial Z-stack projection images were used for quantitative analysis (see Materials and methods). Mean value at each time point is connected by the line to visualize the average intensity change over time. Green, initial values at time point 0; Cyan, mock treatment; Purple, iEPF1 induction. Experiments were repeated three times; n = 6 for time point 0; n = 3 per subsequent time point. Total of 817 nuclei were analyzed. (F) Quantitative analysis of GFP-SCRM levels after iEPF1 induction at the time indicated on 3-day-old seedlings. Each dot represents the total intensity value of GFP signals from each nucleus expressing SCRMpro:: GFP-SCRM. To fully cover the entire nuclei expressing GFP-SCRM, serial Z-stack projection images were used for quantitative analysis (see Materials and methods). Mean value at each time point is connected by the line to visualize the average intensity change over time. Purple, initial values at time point 0; Cyan, mock treatment; Magenta, iEPF1 induction. Experiments were repeated three times; n = 6 for time point 0; n = 3 per subsequent time point. Total of 368 nuclei were analyzed.

تفاصيل العنوان

Figure 4. Induced EPF1 overexpression has no effect in SPCH promoter activity and SPCH protein levels. ; (A) Confocal microscopy images of abaxial cotyledon epidermis from 3-day-old estradiol-inducible EPF1 seedlings expressing SPCHpro::nucGFP (left) and SPCHpro::SPCH-GFP (right) that were either mock treated (top) or treated with estradiol (bottom) from germination for induced EPF1 overexpression (iEPF1). Both SPCH promoter activity and SPCH-GFP protein are robustly detected in arrested meristemoids (arrowheads) by iEPF1 at 3-days post germination (dpg). F1 seedlings were used for the analysis. Scale bars, 20 µm. Experiments were repeated three times. Three seedlings were analyzed each time. (B) Confocal microscopy images of abaxial cotyledon epidermis from 5-day-old estradiol-inducible EPF1 seedlings expressing SPCHpro::nucGFP (left) and SPCHpro::SPCH-GFP (right) that were either mock treated (top) or treated with estradiol (bottom) from germination for induced EPF1 overexpression (iEPF1). At 5-days post germination (dpg), SPCH-GFP proteins have diminished, reflecting the stomatal cell-lineages have past the early stage. F1 seedlings were used for the analysis. Scale bars, 20 µm. Experiments were repeated three times. Three seedlings were analyzed each time. (C) Quantitative analysis of SPCHpro::nucGFP levels after iEPF1 induction at the time indicated on 3-day-old seedlings. Each dot represents the total intensity value of GFP signals from each nucleus expressing SPCHpro::nucGFP. To fully cover the entire nuclei expressing GFP, serial Z-stack projection images were used for quantitative analysis (see Materials and methods). Mean value at each time point is connected by the line to visualize the average intensity change over time. Green, initial values at time point 0; Cyan, mock treatment; Purple, iEPF1 induction. Experiments were repeated three times; n = 6 for time point 0; n = 3 per subsequent time point. Total of 790 nuclei were analyzed. (D) Quantitative analysis of SPCH-GFP levels after iEPF1 induction at the time indicated on 3-day-old seedlings. Each dot represents the total intensity value of GFP signals from each nucleus expressing SPCHpro::SPCH-GFP. To fully cover the entire nuclei expressing SPCH-GFP, serial Z-stack projection images were used for quantitative analysis (see Materials and methods). Mean value at each time point is connected by the line to visualize the average intensity change over time. Purple, initial values at time point 0; Cyan, mock treatment; Magenta, iEPF1 induction. Experiments were repeated three times; n = 6 for time point 0; n = 3 per subsequent time point. Total of 403 nuclei were analyzed.

تفاصيل العنوان

Figure 7—figure supplement 1. Rhodococcus isolates PBTS1 and PBTS2 do not cause disease symptoms in tested plant species. ; (A) Representative images of disease symptoms or beneficial effects on seedlings of N. benthamiana. Three-day-old seedlings were inoculated with Rhodococcus or water (mock) and grown vertically for seven days under constant light. (B) Quantification of seedling root length of N. benthamiana. All were significant relative to mock. There were no significant (ns) differences between treatments with PBTS1, PBTS2, or their mixture. (C, D) Quantification of stem number (C) and stem length (D) of peas inoculated with Rhodococcus or mock-inoculated. * indicates a significant difference when comparing to the mock treatment. (E, F) Quantification of height (E) and internode length (F) of pistachio UCB1 rootstock inoculated with Rhodococcus or mock-inoculated. No treatment effect was significantly different from the time-matched mock treatment. One representative experiment of three is shown.

تفاصيل العنوان

Figure 2—figure supplement 1. The stability of FLS2 clusters depends on the PM lipid composition. ; (A, B) VAEM micrograph of FLS2-GFP (A) and a kymograph of the corresponding VAEM time series (B) after mock treatment. (C, D) VAEM micrograph of BRI1-GFP (C) and a kymograph of the corresponding VAEM time series (D) after mock treatment. (E, F) VAEM micrograph of FLS2-GFP (E) and a kymograph of the corresponding VAEM time series (F) after methyl-β-cyclodextrin (MβCD) treatment. (G, H) VAEM micrograph of BRI1-GFP (G) and a kymograph of the corresponding VAEM time series (H) after MβCD treatment. Application of MβCD, which extracts sterols from the PM, destabilised FLS2-GFP and BRI1-GFP clusters as indicated by the kymographs in (F) and (H). The VAEM micrograph and time series was acquired from a 5 day old Arabidopsis seedling cotyledon after 20 min incubation in 30 mM MβCD, dissolved in liquid MS medium, or mock solution (liquid MS medium). The white arrows in (A), (C), (E) and (G) indicate the spatial dimensions of the kymographs. The scale bars represent 5 µm.

تفاصيل العنوان

Figure 2—figure supplement 3. SE binds and affects the expression strength of CBF genes. ; (A) Visualization of SE ChIP-seq data in WT and se-1. Tracks showing read counts of sequencing reads mapped to the CBF1/2/3 locus. (B) Quantification of indicated RNA transcript level by qPCR in WT and se-3 mutants after 15 or 60 min of cold stress or mock treatment treatment. Error bars denote the standard error of 3 biological replicates.

تفاصيل العنوان

Figure 2—figure supplement 4. The symptoms of Fny-CMV infection in Arabidopsis plants. ; (A) Arabidopsis leaves showing symptoms of viral infection after inoculation with Fny-CMV. Compared to plants treated with water (mock), the leaves of the Fny-CMV-treated plants start to become wrinkled (indicated by white arrows) at day three after inoculation. The edges of leaves are marked with dashed red lines. Fny-CMV inoculation causes obvious distortion of systemically infected small leaves at day 14. Bar = 0.5 cm. (B) Images of Arabidopsis leaves after inoculation with Fny-CMV or mock treatment (water). All leaves from Arabidopsis plants 14 days after treatment are presented. Bar = 1 cm. (C) Quantification of average leaf length in WT Arabidopsis plants 2 weeks after inoculation with Fny-CMV. The long axis of the blade (the red arrow) was measured as the leaf length. Values represent mean ± SE, n ≥ 20. The experiments were repeated at least five times. (D) Quantification of average Arabidopsis leaf length after inoculation with Fny-CMV at days 12, 13 and 14. Values represent mean ± SE, n ≥ 20. **p 3.

تفاصيل العنوان

Figure 3—figure supplement 2. The stability of RNAs produced by SE target genes is not affected SE. ; Analysis of RNA stability in WT, se-1 and se-3 mutants. Plants grown in liquid culture were treated with 200 µg/ml cordycepin for 0.5 hr, 1 hr and 2 hr to block transcription. Mock treatment served as a control. Quantification of indicated RNA transcript level was performed by qPCR. Error bars denote the standard error of 3 biological replicates.

تفاصيل العنوان

Figure 1—figure supplement 2. Subcellular localisation (A, B) and co-immunoprecipitation analysis (C) of pKNOLLE::mRFP:PEP12 (aka SYP21) (red) in big3 mutant seedling root cells expressing estradiol-inducible YFP:NPSN11 (A, C) and GFP:SNAP33 (B, C). ; Transgenic seedlings were treated with 50 µM BFA for 30 min followed by 50 µM BFA + 20 µM estradiol for 210 min. Protein extracts were subjected to immunoprecipitation with anti-GFP beads, protein blots were probed with the antisera indicated on the right (IB): GFP, anti-GFP; RFP, anti-RFP; KN, anti-KNOLLE; V721/V722, anti-VAMP721/722; SYP71, anti-SYP71. G:S33, GFP:SNAP33; R:P12, mRFP:PEP12; Y:N11, YFP:NPSN11. kDa, protein size (left); MW, molecular weight; -BFA, mock treatment; +BFA, BFA treatment; T, total extract; UB, unbound; IP, immunoprecipitate. Loading (%), relative loading volume to total volume; relative signal intensity (input signal = 100% for UB and IP). Nuclei of merged images (A, B) were counterstained with DAPI (blue). Arrowheads, cell plates; arrows, BFA compartments. Scale bar, 5 µm. The experiments were technically repeated three times.

تفاصيل العنوان