The role of mGluR2 versus mGluR3 in the ischemic control of Cx36 expression. Data from siRNA (A, B) and Western blot (A–D) experiments in wild-type mouse neuronal cortical cultures are shown. A, B, Transfections were done on day in vitro 7 (DIV7), followed by OGD or sham OGD on DIV9 and Western blot analysis 2 h after OGD/sham OGD. In sham OGD conditions (control), siRNA suppression of mGluR2 (A) and mGluR3 (B) decreases the receptor protein levels but does not significantly affect background levels of Cx36. Following OGD, the cells transfected with scrambled siRNA (SCR siRNA) demonstrate increased Cx36 expression, but this increase does not occur in cells transfected with active siRNA (either mGluR2 or mGluR3). C, Developmental changes in the expression of mGluR2 and mGluR3. D, In mature cultures, BINA (3 μm; 50 min) potentiates the effect of a 5-min-long OGD on Cx36 expression. In all figures, statistical data and representative blots (a1–d1) are shown. Optical density signals are normalized relative to tubulin, and normalized values are compared with the scrambled siRNA in control conditions (A, B), corresponding DIV7 (C), and control (D), set at 1.0. Statistical analysis was as follows: ANOVA with post hoc Tukey (A, B, D); Student's t test (C); *p < 0.05, **p < 0.01, ***p < 0.001; n = 5–10 (A, B), 4 (C), 3 (D) per group; data are shown as mean ± SEM. Stainings were done sequentially on one membrane.