To test whether USP35 affect the expression of ERα target genes, mRNA levels of pS2, GREB1, Myc, and CCND1 were analyzed in ER+ breast cancer cells with USP35 overexpression (Fig. 5a) or USP35 knockdown (Fig. 5b) by qRT-PCR. The gene discussed is CCND1; the disease is breast cancer.