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Fig. 5. Effects of MRTF-A knockout on phenotypic changes. Cells at ~70% confluence were transfected with control plasmids (A, lane 1) or the MRTF-A gene was knocked out using MRTF-A HDR CRISPR-associated 9 (Cas9) (A, lane 2) for three days. On day 2 postconfluence, the expression of MRTF-A, N-cad, sm MHC, nm MHC, and PP1β in the supernatant fraction (80 μg) was determined as described in the legend of . p-values were obtained by unpaired Student’s t-tests. (B) Control and knockout cells were incubated before and after stimulation with ML-7 (6 × 10−5 M) for 1 h each, and secreted active renin was measured by ELISA (B, n = 4). MRTF-A, myocardin related transcription factor-A; HDR, homology directed repair; CRISPR, clustered regularly interspaced short palindromic repeats; sm MHC, smooth muscle myosin heavy chain; nm MHC, nonmuscle myosin heavy chain; PP1β, protein phosphatase 1β. NS, no significant difference. p-values were obtained either by paired Student’s t-tests (within groups) or by ANOVA (between groups).
Korean J Physiol Pharmacol 2022;26:479-499
© Korean J Physiol Pharmacol