Fig. 2: MTHFD2 knockdown inhibits proliferation, induces apoptosis and causes G0/G1 arrest in vitro.

A Western blot assay showing the MTHFD2 and actin protein level of shNC (Negative control), shMTHFD2#1, shMTHFD2#2 in NCI-H929 and OPM2 cells. B CCK8 assay was used to quantitatively detect the proliferation of MTHFD2 knockdown NCI-H929 and OPM2 cells using OD values at the corresponding time. Data were presented as mean ± SD from three independent experiments. C Annexin V /PI double staining flow cytometry was used to detect the apoptotic rate after knockdown of MTHFD2 in NCI-H929 and OPM2 cells. D Results as bar graph are calculated percentages of Annexin-V positive cell populations from three independent experiments. E The apoptosis related proteins including PARP1, cleaved-PARP1, caspase3, cleaved-caspase3, and internal reference actin were detected by western blot assay in MTHFD2 knockdown NCI-H929 and OPM2 cells. F Cell cycle of NCI-H929 and OPM2 cells was examined after knockdown of MTHFD2.Results were presented as peak plots using Modfit. G The distributions of cells in G0/G1, S and G2/M phases were calculated. Data represent mean ± SD from triplicate independent experiments. H Western blot showed protein expression of cyclin D1 and CDK4 in MTHFD2-silenced NCI-H929 and OPM2 cells. The data are presented as mean ± SD. Student’s t test was used to compare the differences between two groups. (***P < 0.001, ****P < 0.0001).