Figure 4.

Knockdown of GPR30 depressed estrogen-induced neuroprotection. Cells were treated with GPR30-shRNA for 24 h. A, Confocal images showing the transfection rate of the GPR30-shRNA. Top, Nuclei were stained with Hoechst33258 (blue). GFP-positive neurons indicated the successful knockdown of GPR30 by transfection with GPR30-shRNA. Bottom, Typical neurons showing GPR30-shRNA transfection (green) and nontransfection (red). Scale bars: 20 μm. B, Transfection of GPR30-shRNA resulted in a reduction in GPR30 protein levels; the negative shRNA did not lead to the change of GPR30 expression. ^$$p < 0.01 compared with the control and negative shRNA. C, Transfection of GPR30-shRNA blocked the neuroprotection of G1 and slightly inhibited the effects of E2. ^$p < 0.05 compared with G1 treated negative shRNA; *p < 0.05, **p < 0.01 compared with NMDA treatment alone.