Fig. 5.

Decreased Tns2 mRNA expression results in increased formation of actin stress fibers. (A) Real-time PCR analysis of Tns2 mRNA. Three days after siRNA transfection, Tns2 mRNA expression significantly decreased in Tns2 KD cells (n=3). (B) Adhesion assay of Tns2 KD cells. Cells were seeded on culture dishes coated with collagen type IV (C), fibronectin (F), laminin (L) or vitronectin (V). After incubation for 1 hr, non-adherent cells were removed by gentle washing with PBS and then counted (n=3). (C)–(D) Quantification of phalloidin staining in Tns2 KD cells. The degree of actin stress fiber formation was classified into three categories (high to low: a, b and c), depending on the thickness and length of the actin stress fibers in the cytoplasm. In Tns2 KD podocytes, cells in the stress fiber-rich category, with thick cables, (type a) increased, whereas cells lacking stress fibers, without thick cables, (type c) decreased, suggesting that Tns2 suppression significantly enhanced actin stress fiber formation (Fig. 5C and 5D). Tns2-KD podocytes were compared with control cells in 5 separate fields (n=5).