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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: FEBS J. 2018 May 24;285(15):2785–2798. doi: 10.1111/febs.14498

Figure 6. Cellular apoptosis is induced through pSTAT6 by reduction of IL-4Rα expression level using Il4rα-specific siRNA in the cortical brain slice ischemia assay.

Figure 6

(A) Non-specific (control) or Il4rα-specific siRNA were transfected into Il4rα WT cortical brain slices for 48 h. mRNA levels of Il4, Il4rα from brain slices transfected either control or Il4rα-specific siRNA were determined by qRT-PCR 24 h after 30 min OD. Gapdh was used for normalization. Experiments were performed three times using three mice per genotype for each experiment. (B) Western blots were performed to detect IL-4Rα in explanted brain slices of Il4rα WT brain slices transfected with either control siRNA or Il4rα-specific siRNA in both control and OD conditions. (C) Levels of IL-4Rα protein were normalizing to GAPDH protein levels. Experiments were performed three times using three mice per genotype for each experiment. Data represent the mean ± SEM. Statistical analysis, 2-tailed Student’s t test (* p<.05 vs. non-specific siRNA). (D) Non-specific (control) or Il4rα-specific siRNA were transfected into wild-type cortical brain slices. 48 h later mRNA levels of Il4, Il4rα, Stat6, Bcl-2, and Caspase 3 from Il4rα WT were determined by qRT-PCR 24 h after 30 min OD. Gapdh was used for normalization. Experiments were performed three times using three mice per genotype for each experiment. (E) Western blots were performed to detect both pSTAT6 and STAT6 in explanted brain slices of Il4rα WT mice transfected with either control siRNA or Il4rα-specific siRNA for both control and OD conditions. (F) Levels of pSTAT6 protein were determined using total STAT6 protein levels for normalization. (G) Western blot were performed to detect both BCL-2 and c-Caspase 3 in explanted brain slices of Il4rα WT and KO transfected with either control siRNA or Il4rα-specific siRNA for both control and OD conditions. (H and I) Levels of BCL-2 (H) and c-Caspase 3 (I) proteins were determined using normalizing to GAPDH protein levels. Experiments were performed four times using three mice per genotype for each experiment. Data represent the mean ± SEM. Statistical analysis, two-way ANOVA followed by Bonferroni’s multiple comparison test (* p<.05; ** p<.01; *** p<.001).