Journal: Journal of Cellular and Molecular Medicine
Article Title: Essential role of Pin1 via STAT3 signalling and mitochondria-dependent pathways in restenosis in type 2 diabetes
Figure Lengend Snippet: Effects of Pin1 on proliferation, cell cycle progression, apoptosis and migration in vascular smooth muscle cells (VSMCs). VSMCs were cultured and infected as described in Figure 3 . Cell proliferation was assessed by both the MTT assay ( A ) and [ 3 H]-thymidine incorporation analyses ( B ). Cell cycle analysis was quantified by PI staining followed by flow cytometry analyses. M1, M2, M3 and M4 indicate sub G1, G0/G1, S and G2/M phases respectively ( C ). Bar graphs represent the mean ± SEM of three independent experiments ( D ). Cell apoptosis was detected by annexin V-FITC/PI staining ( E ) followed by flow cytometry analysis and TUNEL staining ( G ). The proportion of live cells (annexin V−/PI−), early apoptotic cells (annexin V+/PI−), late apoptotic/necrotic cells (annexin V+/PI+) and dead cells (annexin V−/PI+) was calculated for comparison ( F ). The TUNEL-positive cells were stained green, VSMCs were stained for SMαA and shown in red. Nuclei were stained blue by DAPI. The percentages of apoptotic nuclei were calculated by determining the number of DAPI-stained nuclei that were also positive for TUNEL staining. Approximately, 100 nuclei cells were counted in randomly chosen fields per region ( H ). VSMC migration was determined by a standard wound healing assay ( I ). Bar graphs represent the mean ± SEM of three independent experiments ( J ). * P
Article Snippet: Thereafter, cells were incubated with anti-α-actin and anti-pin1 antibody (Abcam, Cambridge, UK) and further stained with appropriate TRITC- or FITC-conjugated secondary antibodies (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).
Techniques: Migration, Cell Culture, Infection, MTT Assay, Cell Cycle Assay, Staining, Flow Cytometry, Cytometry, TUNEL Assay, Wound Healing Assay