Fig. 8

Rasal1 promotes CaMKII phosphorylation. A Representative confocal images of blocked control hippocampal neurons treated with TTX, CNQX and APV for at least 8 h to block CaMKII phosphorylation (A1), EGFP-control hippocampal neurons treated for 5 min with 10 µM glycine to acutely induce CaMKII phosphorylation (A2), glycine (10 µM) treated Rasal1-OE transduced hippocampal neurons (A3) and glycine (10 µM) treated Rasal1-shRNA transduced hippocampal neurons (A4) stained for phosphorylated (red) or total CaMKII (cyan). Transfected neurons (green) indicated by arrows were compared to neighboring non-transfected neurons. Scale bar: 20 µm. B Quantification of the ratio of phosphorylated CaMKII fluorescence intensity of transfected /non-transfected EGFP-control (n = 20), Rasal1-OE (n = 25), and Rasal1-shRNA (n = 25) transduced hippocampal neurons. C Quantification of total CaMKII levels in EGFP-control (n = 20), Rasal1-OE (n = 25), and Rasal1-shRNA (n = 25) transduced hippocampal neurons as ratio of fluorescence intensity in transfected/non-transfected neurons. D Fluorescent western blot of p-CaMKII (red) of glycine (10 µM) treated EGFP-control, Rasal1-OE, and Rasal1-shRNA transduced hippocampal neurons. E Fluorescent western blot of CaMKII (green) and p-CaMKII (red) of glycine (10 µM) treated EGFP-control, Rasal1-OE, and Rasal1-shRNA transduced hippocampal neurons. F Quantification of p-CaMKII/actin levels in fluorescent western blot across of glycine (10 µM) treated EGFP-control, Rasal1-OE, and Rasal1-shRNA transduced hippocampal neurons (n = 8). G Quantification of total CaMKII/actin levels in fluorescent western blot across of glycine (10 µM) treated EGFP-control, Rasal1-OE, and Rasal1-shRNA transduced hippocampal neurons. (n = 8). Data information: In (B, C, F, G) *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed unpaired student t-test