Figure 3

GPER mediates the promotion effect of E2 on HOTAIR expression. A): MDA-MB-231 cells were treated with 1 μM G1 with or without 100 nM G15 for 6 h. Then the expression of the p-ERK level was checked with a western blot. B): MDA-MB-231 and BT549 cells were pretreated with 100 nM G15 for 6 h before the addition of 10 nM E2 for 24 h. Then the expression of HOTAIR was determined by real-time PCR. C): The HOTAIR promoter sequence was cloned into PGL3-basic luciferase reporter plasmid, and then the control plasmid and HOTAIR promoter including plasmid were transfected into the MDA-MB-231 and BT549 cells. Then 10 nM E2 was added into these transfected cells. After E2 treatment for 24 h, the luciferase activity was detected. The results are shown as mean ± S.E. from three representative independent experiments. **P < 0.01 compared with control. ^#p < 0.05 compared with E2. ns p > 0.5.