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. 2019 Feb 6;316(5):C621–C631. doi: 10.1152/ajpcell.00208.2018

Fig. 5.

Fig. 5.

Polymerase-δ-interacting protein 2 (Poldip2) and epithelial cell transforming sequence 2 (Ect2) both regulate VSMC cytokinesis and proliferation. A–C: rat aortic smooth muscle cells (RASMs) were plated on collagen-coated coverslips and transfected with either of 2 different siRNA sequences against the same target: control (siCtl-1 and 2), Poldip2 (siPoldip2-A and B), or Ect2 (siEct2-A and B). 72 h after siRNA transfection, the cells were lysed to confirm knockdown by immunoblot (A) or stained with phalloidin (green), wheat germ agglutinin (red), and DAPI (blue) (B). The number of multinucleated cells (examples indicated by white arrow) in each condition was counted by a blinded observer and the average percentage of multinucleated cells graphed (n = 4–7; ****P < 0.0001 vs. siCtl-1; ####P < 0.0001 vs. siCtl-2 Cochran-Mantel-Haenzel test; error bars = SE) (C). D: a separate group of RASMs was plated in 6-well plates in equal numbers. The following day, the cells were treated in quintuplicate with control (siCtl-2), Poldip2, or Ect2 siRNA. Knockdown was confirmed by immunoblot at 24 h after transfection and the number of RASMs in each condition was counted every 24 h for 5 days following transfection at time 0 (n = 3; ####P < 0.0001 vs. siCtl-2 at 72-, 96-, and 120-h two-way ANOVA with Tukey’s method for correction; error bars represent SE).