Fig. 4.

Knockdown of SHMT2 inhibited the proliferation and modulates the TSCC’s cell cycle in vitro. a, b Western blot analysis and real time PCR of HN6 and HSC3 cells transfected with SHMT2 siRNA. c, d CCK8 assay performed on HN6 and HSC3 cells after transfection to determine the cell viability. e, f Representative images of and quantification of EdU stained cells in negative control and SHMT2 siRNA transfected groups. g, h Flow cytometric analysis of indicated cells, representative graph (left) and quantitative analysis (right). i Western blot analysis of several cell cycle regulators including P21, P27, CDK4, and cyclin D1 expression in indicated cells. j Real time PCR analysis of several cell cycle regulators including CDKN1A, CDKN1B, CCDN1 and CDK4 in HN6 and HSC3. P-values were obtained using Student's t-tests and two-way ANOVA tests * p < 0.05, ** p < 0.01, *** p < 0.001