Fig. 3

Roquin1 selectively inhibited the mRNA expression of cell cycle–promoting genes via targeting the 3′UTR. a RNA-seq analysis showed changes in the expression levels of cell cycle–related genes by Roquin1 in MCF7 cells. The genes promoting G1/S, S phase, G2/M, and M phase transition were downregulated; and the expression of cell cycle progression inhibitor p21 was upregulated. b Top 10 KEGG pathways enriched for common downregulated genes of Roquin1. c Top 10 GO terms (biological processes) for the analysis for common downregulated genes. d-e The mRNA levels of indicated cell cycle–promoting genes were measured by qPCR in MCF7 (d) and MDA-MB-468 (e) cells at indicated time points. *P < 0.05; **P < 0.01 between two groups. f-g CCNE1 and MCM2 were measured by immunoblotting with anti-CCNE1 and anti-MCM2 antibodies at different time points after Roquin1 overexpression in MCF7 (f) and MDA-MB-468 (g) cells. h-i Roquin1/GFP fusion protein were expressed in MDA-MB-468 cells, after lysate extraction, immunoprecipitation with anti-GFP antibody or IgG, and RNA extraction. Cell cycle–promoting gene (h) and –inhibiting gene (i) transcripts were detected by RT-PCR. j Measurement of luciferase activity of reporters containing the 3′UTRs of CCND1, CCNE1, CKD6 (part), and MCM2, respectively. β-Actin 3′UTR was used as a negative control. The results shown represent the mean ± standard deviation of four independent experiments. *P < 0.05