sm22α (tagln) rabbit pab, a6760 antibody  (ABclonal Biotechnology)

Journal: Biomedicines

Article Title: RhoGDI1-Cdc42 Signaling Is Required for PDGF-BB-Induced Phenotypic Transformation of Vascular Smooth Muscle Cells and Neointima Formation

doi: 10.3390/biomedicines9091169

Figure Lengend Snippet: MG132 pretreatment blocks the reduced phenotypic transition of HA-VSMCs induced by crenolanib. HA-VSMCs were pretreated with crenolanib (50 nM) for 48 h or MG132 (5 μM) for 1 h and then exposed to 10 ng/mL PDGF-BB treatment for 24 h. Untreated cells were used as control. ( A ) Cell proliferation was checked by EdU assay (scale bar = 100 μm). Histogram showing the ratio of EdU-positive cells (red) to the total number of cells. **, p < 0.01 vs. the control group; ##, p < 0.01 vs. the group treated with PDGF-BB; $$, p < 0.01 vs. the PDGF-BB and crenolanib-treated group ( n = 3). ( B ) Cell migration was checked by wound-healing (scale bar = 200 μm) and transwell assays (scale bar = 100 μm). Histograms showing the quantification of the wound healing (migrating cells from the scratched boundary) and transwell assay (crystal violet stained cells migrating to the lower chamber) results. **, p < 0.01 vs. the control group; #, p < 0.05 and ##, p < 0.01 vs. the PDGF-BB-treated group; $, p < 0.05 vs. the PDGF-BB and crenolanib-treated group ( n = 3). These results show that MG132 pretreatment promoted the reduced cell proliferation and migration mediated by crenolanib. ( C ) Western blot analysis of MYH-11, SM22, and smoothelin. MG132 reduced the increased smoothelin expression induced by crenolanib. Histograms showing the ratios of target proteins to β-tubulin. *, p < 0.05 vs. the control group; #, p < 0.05 vs. the PDGF-BB-treated group; $, p < 0.05 vs. the PDGF-BB and crenolanib-treated group ( n = 3).

Article Snippet: Anti-RhoGDI1 (A1214), –Ubiquitin (A3207), –smoothelin (# 6745), –MYH11 (# 10827), -ki-67 (A11390), and -SM22α (# A6760) antibodies were purchased from ABclonal (Wuhan, China).

Techniques: Control, EdU Assay, Migration, Transwell Assay, Staining, Western Blot, Expressing

Journal: Biomedicines

Article Title: RhoGDI1-Cdc42 Signaling Is Required for PDGF-BB-Induced Phenotypic Transformation of Vascular Smooth Muscle Cells and Neointima Formation

doi: 10.3390/biomedicines9091169

Figure Lengend Snippet: RhoGDI1 suppression inhibits PDGF-BB-induced phenotypic transition in HA-VSMCs. Cells were transfected with RhoGDI1 siRNA for 48 h and then treated with 10 ng/mL of PDGF-BB for another 24 h. Untreated cells were used as control. ( A ) Confirmation of RhoGDI1 knockdown by western blotting. ( B ) EdU assay showed that RhoGDI1 suppression inhibited PDGF-BB-induced cell proliferation (scale bar = 100 μm). Histogram showing the ratio of EdU-positive cells (red) to total cells. **, p < 0.01 vs. the control group; ##, p < 0.01 vs. the group treated with PDGF-BB ( n = 3). ( C ) Wound-healing (scale bar = 200 μm) and transwell assays (scale bar = 100 μm) showed that RhoGDI1 knockdown reduced cell migration induced by PDGF-BB. The quantification method refers to . **, p < 0.01 vs. the control group; #, p < 0.05 vs. the PDGF-BB-treated group ( n = 3). ( D ) Western blot analysis of MYH-11, SM22, and smoothelin showed that RhoGDI1 suppression promoted the expression of smoothelin in PDGF-BB-treated cells. Histograms showing the ratios of target proteins to β-tubulin. *, p < 0.05 vs. the control group; #, p < 0.05 vs. the PDGF-BB-treated group ( n = 3).

Article Snippet: Anti-RhoGDI1 (A1214), –Ubiquitin (A3207), –smoothelin (# 6745), –MYH11 (# 10827), -ki-67 (A11390), and -SM22α (# A6760) antibodies were purchased from ABclonal (Wuhan, China).

Techniques: Transfection, Control, Knockdown, Western Blot, EdU Assay, Migration, Expressing

Journal: Biomedicines

Article Title: RhoGDI1-Cdc42 Signaling Is Required for PDGF-BB-Induced Phenotypic Transformation of Vascular Smooth Muscle Cells and Neointima Formation

doi: 10.3390/biomedicines9091169

Figure Lengend Snippet: Cdc42 inhibition reduces PDGF-BB-induced phenotypic transition in HA-VSMCs. HA-VSMCs were pretreated with ZCL278 (50 μM) for 30 min followed by treatment with 10 ng/mL of PDGF-BB for 24 h. Untreated cells were used as control. ( A ) Cell proliferation was analyzed by EdU assay (scale bar = 100 μm). Histogram showing the ratio of EdU-positive cells (red) to total cells. **, p < 0.01 vs. the control group; #, p < 0.05 vs. the PDGF-BB-treated group ( n = 3). ( B ) Cell migration was analyzed by wound-healing (scale bar = 200 μm) and transwell assays (scale bar = 100 μm). The quantification method refers to . **, p < 0.01 vs. the control group; #, p < 0.05 vs. the PDGF-BB-treated group ( n = 3). ZCL278 pretreatment significantly decreased cell proliferation and migration induced by PDGF-BB. ( C ) Western blot analysis of MYH-11, SM22, and smoothelin. ZCL278 pretreatment increased the expression of smoothelin in PDGF-BB-treated cells. Histograms showing the ratios of the target proteins to β-tubulin. *, p < 0.05 vs. the control group; #, p < 0.05 vs. the PDGF-BB-treated group ( n = 3).

Article Snippet: Anti-RhoGDI1 (A1214), –Ubiquitin (A3207), –smoothelin (# 6745), –MYH11 (# 10827), -ki-67 (A11390), and -SM22α (# A6760) antibodies were purchased from ABclonal (Wuhan, China).

Techniques: Inhibition, Control, EdU Assay, Migration, Western Blot, Expressing

Journal: Biomedicines

Article Title: RhoGDI1-Cdc42 Signaling Is Required for PDGF-BB-Induced Phenotypic Transformation of Vascular Smooth Muscle Cells and Neointima Formation

doi: 10.3390/biomedicines9091169

Figure Lengend Snippet: Both RhoGDI1 and Cdc42 suppression inhibit neointima formation and improve the expression of contractile proteins in a rat carotid injury model. For shRNA-mediated knockdown, the carotid artery was injected with approximately 0.2 mL of virus solution (titer: 1 × 10 10 pfu) after balloon injury operation. ( A ) H&E of arteries 14 days after balloon injury. Rats without balloon injury were used as the sham operation group. Histogram showing the area ratio of intima to media. Both RhoGDI1 and Cdc42 knockdown reduced intima/media area ratio. **, p < 0.01 vs. the sham operation group; ##, p < 0.01 vs. the injury model group ( n = 10). ( B ) Immunofluorescence staining for contractile proteins such as MYH-11, SM22, smoothelin (green), ki-67 (green), and α-SMA (green) and CD31 (red) as double staining (merge); nuclei were stained with DAPI (blue). Histograms showing the fluorescence intensity of the staining. RhoGDI1 and Cdc42 knockdown reduced the expression of ki-67 and co-localization of α-SMA with CD31 and increased the expression of the three contractile proteins. *, p < 0.05 vs. the sham operation group; #, p < 0.05 vs. the injury model group ( n = 10). ( C ) Western blots showing the expression of MYH-11, SM22, and smoothelin. Histograms showing the ratios of the contractile proteins to β-tubulin. The results are consistent with those from immunofluorescence analysis. *, p < 0.05 vs. the sham operation group; #, p < 0.05 and ##, p < 0.01 vs. the injury model group ( n = 10).

Article Snippet: Anti-RhoGDI1 (A1214), –Ubiquitin (A3207), –smoothelin (# 6745), –MYH11 (# 10827), -ki-67 (A11390), and -SM22α (# A6760) antibodies were purchased from ABclonal (Wuhan, China).

Techniques: Expressing, shRNA, Knockdown, Injection, Virus, Immunofluorescence, Staining, Double Staining, Fluorescence, Western Blot