FIG. 3.
MT1H, MT1F, and BRCA1 expression is downregulated in HeLa cells following induction of a PLU-1/JARID1B transgene. (A) Western blot showing PLU-1/JARID1B expression from an inducible promoter. Lysates from HeLa cells transfected with the PLU-1/JARID1B-inducible vector (HeLa-PLU-1-Trex) or from cells transfected with the empty vector (HeLa empty vector) were prepared after inducing the expression of the transgene for 0, 24, and 72 h. The extracts were separated on a denaturing gel, and Western blot analysis was performed with α-PLU-1-C antiserum or anti-HSC70 antibody for loading control. (B) q-RT-PCR analysis of expression of PLU-1/JARID1B, MT1H, MT1F, and BRCA1 in the PLU-1/JARID1B transfected HeLa cells induced for 24 or 72 h. Data for each gene at 24- or 72-h time points are normalized against the internal expression of β-actin and are represented as the change (2-ΔΔCT) compared to the noninduced cells. Bars represent the means ± standard errors of the means of three independent experiments. The asterisks denote a significant difference (P < 0.05) between induced and uninduced cells by using Student's t test.