Fig 1: Inhibition of RhoA/ROCK1 pathway attenuated compression stress-induced human NP cells senescence. (A, B) The level of senescent cells in different groups was assessed by SA-ß-gal staining. Scale bar: 50 µm. (C, D) Human NP cells untreated or pretreated with 20 µM Y27632 for 2 h prior to compression stress for 36 h were stained with MRTF-A (green) and DAPI (blue) (n = 20). Scale bar: 10 µm. (E, F) mRNA levels of the ECM remodelling proteinases (MMP3, MMP13 and ADAMTS5) and ECM component (COL1A, COL2A and aggrecan) were measured after the human NP cells were treated withY27632. (G, H) The proportions of cells in each cycle were measured through flow cytometry in different groups. (I, J) CDK4, Cyclin D1, p21 and p53 expression levels in different groups were measured through Western blot analysis and normalized to that of GAPDH. Data were presented as the mean ± SD (n = 3). #No significance, vs compression group; *P < .05, vs compression group; **P < .01, vs compression group; ***P < .001, vs compression group
Fig 2: The effect of MRTF-A, HDAC5, and p300 on apoptosis of cortical neurons induced by H2O2. Apoptosis was detected by Annexin-V+PI double staining (a and b) and caspase-3 expression (c). (1) control, (2) Vector+H2O2 (400 µM, 24 h), (3) H2O2 (400 µM, 24 h)+MRTF-A, (4) H2O2 (400 µM, 24 h) +p300, (5) H2O2 (400 µM, 24 h)+HDAC5, (6) H2O2 (400 µM, 24 h)+MRTF-A+p300, (7) H2O2 (400 µM, 24 h)+MRTF-A+HDAC5. +++P<0.001 versus control; *P<0.05, ***P<0.001 versus H2O2+vector; #P<0.05, ##P<0.01 versus H2O2+MRTF-A. n=4. Data are representative of four independent experiments
Fig 3: Schematic graph of the role of myosin IIA and IIB in compression stress-induced senescence of NP cells. Compression stress induced the RhoA/ROCK1 pathway activation, which regulated the interaction of myosin IIA and IIB with actin. The actomyosin cytoskeleton remodelling was involved in the compression stress-induced fibrotic phenotype mediated by MRTF-A nuclear translocation and inhibition of proliferation in human NP cells
Fig 4: Expression of MRTF-A, HDAC5, and p300 and their localization in rat brain sections. (a) Representative images showing the expression and localization of MRTF-A and HDAC5 in sections from control rats and from rats after I/R (400 ×). (b) Representative images showing the expression and localization of MRTF-A and p300 in sections from control rats and from rats after I/R. (c) The interaction of MRTF-A and p300 by co-immunoprecipitation. Representative images were from six rats of each group. n=3. Data are representative of three independent experiments
Fig 5: The involvement of MRTF-A/Bcl-2/Mcl-1 downregulated in cerebral ischemia/reperfusion model. MRTF-A (a), Bcl-2 (b), and Mcl-1 (c) protein expression was downregulated. LV-MRTF-A-siRNA was transfected in the brain tissue (d, 100 ×). MRTF-A protein expression was detected (e). The apoptosis of brain neurons was detected by TUNEL (f–g, 400 ×) and cleaved caspased-3 (h). *P<0.05, **P<0.01, ***P<0.001 versus sham; ++P<0.001 versus LV-N+I/R 24 h. n=5–6 in each group. LV-N: lentivirus-negative-siRNA; LV-MRTF-A-siRNA: lentivirus-negative-MRTF-A-siRNA
Supplier Page from Abcam for Anti-Mkl1/MRTFA antibody