Fig. 7
Changes in AJ gene expression and F-actin arrangement in floating hHFMSCsOCT4 after ZO-1 knockdown. (a) RT‒qPCR and Western blot showing the expression of ZO-1 after RNAi (n = 2). Full-length blots/gels are presented in Supplementary Figure S4. (b) RT‒qPCR and detection of the mRNA expression of ACTN2, E-cadherin, β-catenin, and Cyclin D1 (n = 3). The error bars represent the standard deviations of measurements in three separate sample runs. (c) Western blots showing the protein expression levels of β-catenin (n = 1). Full-length blots/gels are presented in Supplementary Figure S5, n = 1. (d) Phalloidin staining showing F-actin morphology and distribution (n = 3). (e and f) Fluorescence microscopy (Olympus) captured DAPI-stained cell images, and CellProfiler software detected nuclei and calculated their integrated intensity. R software then used visually determined cutoffs to calculate the percentages of cells in G1, S, and G2/M phases. NC, negative control. si-ZO-1, ZO-1-knockdown. *P˂0.05, **P˂0.01, ***P˂0.001, ****P˂0.0001