Fig. 5: Gαi1 and Gαi3 are required for RSPO3-induced Erk activation and neuroprotection against OGD/R. | Cell Death & Disease

Fig. 5: Gαi1 and Gαi3 are required for RSPO3-induced Erk activation and neuroprotection against OGD/R.

From: Endothelial cell-derived RSPO3 activates Gαi1/3-Erk signaling and protects neurons from ischemia/reperfusion injury

Fig. 5

Wild-type (WT), Gαi1/3 double knockout (DKO), Gαi1, Gαi2 or Gαi3 single knockout (SKO) mouse embryonic fibroblasts (MEFs) were treated with RSPO3 (at 50 ng/mL) and cultivated for indicated time periods, expression of listed proteins was shown (A, B). Neuro-2a cells, with the Gαi1 shRNA plus the Gαi3 shRNA (“Gαi1/3-DshRNA”), the scramble control shRNA (“scr-shRNA”), were established and expression of listed genes and proteins was shown (C, D); Cells were treated with RSPO3 (50 ng/mL) or vehicle control and cultured for 15 min, expression of listed proteins was tested (E). Gαi1/3-DshRNA or scr-shRNA Neuro-2a cells were pretreated with RSPO3 (50 ng/mL) for 30 min, cells were then maintained under oxygen glucose deprivation (OGD) for 4 h and then re-oxygenation (“OGD/R”) for the applied time periods, cell viability, death and apoptosis were tested by CCK-8 (F), medium LDH release (G) and nuclear TUNEL staining (H) assays, respectively. “Mock” stands for the mock treatment (norm-oxygenated medium with glucose). Data were presented as mean ± standard deviation (SD, n = 5). *P < 0.001 vs. “WT” MEFs/“scr-shRNA”. #P < 0.001. Each experiment was repeated five times and similar results were obtained.

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