Fig. 4.

Impact of FOXO3 and YTHDF2 Knockdown and overexpression on the expression of TIMP1 and MMPs. (A) mRNA expression levels of YTHDF2, TIMP1, and MMPs in FOXO3^KD and FOXO3^OE cells. Quantitative PCR was performed on RNA from control knockdown (Control^KD-1 and Control^KD-2), FOXO3 knockdown (FOXO3^KD-1 and FOXO3^KD-2), control overexpression (Control^OE-1 and Control^OE-2), and FOXO3 overexpression (FOXO3^OE-1 and FOXO3^OE-2) cell lines (NP background) to assess levels of FOXO3, YTHDF2, TIMP1, MMP1, MMP3, MMP7, and MMP9. (B) mRNA expression levels of FOXO3, TIMP1, and MMPs in YTHDF2^KD and YTHDF2^OE cells. RNA from Control^KD-1/2, YTHDF2^KD-1/2), Control^OE-1/2, and YTHDF2^OE-1/2 cell lines (NP background) was analyzed to detect the expression levels of the same genes as in (A) by RT-qPCR. (C) Protein expression of YTHDF2, TIMP1, and MMPs in FOXO3^KD and FOXO3^OE cells. Protein samples from cells in (A) were analyzed by Western blot to detect levels of the indicated proteins. (D) Protein expression of FOXO3, TIMT1, and MMPs in YTHDF2^KD and YTHDF^OE cells. Protein samples from cells in (B) were subjected to Western blot analysis to detect levels of the indicated proteins. (E) The binding of FOXO3 on the promoter of TIMP1 by ChIP assay using cells in (A). (F) The binding of YTHDF2 on the promoter of TIMP1 by ChIP assay using cells in (B). **P < 0.01, ***P < 0.001