Figure 3.

Confirmation of overexpressed SPRY1 protein in the transgenic mice. A, Genotyping was performed by PCR products electrophoresis. The left line was DNA markers, 330 bp band was SPRY1 and Rsg7 (632 bp) was selected as internal control. Western blot analysis of tissue samples from epidermis of SPRY1‐TG and WT. β‐actin was used as internal control. B, SPRY1 expression by immunofluorescence and immunochemistry staining of mice skin from SPRY1‐TG and WT mice respectively. The tissue sections were also stained with DAPI to visualize the nuclei. Scale bar = 100 μm for all panels. C, qRT‐PCR analysis relative mRNA expression of cell cycle‐associated genes (p21, p27, cyclinb1, g0s2, p53) in SPRY1‐TG and WT mice skin. Vertical axis indicated relative expression of mRNA by GAPDH as control. n.s: no statistically significant. ***P < .001. Symbols indicate mean mean ± SEM of six mice per group with student t test. D,E, Images of western blot showed alteration of cell cycle‐related proteins (P27, P21, p‐P53), EMT‐related proteins (K1/K10, β‐catenin, MMP3 and K14 and caspase‐3. β‐actin was used as internal control (Lysate was obtained from epidermis of SPRY1 TG and wild‐type mice)