Fig. 6. AQP4 knockout downregulates the expression of Cav1.2 and connexin43 in ANSCs. (A) RT-PCR analysis shows Cav1.2, Cav3.1 and TRPC1 mRNA is present in the ANSCs. The transcripts of Cav1.3, Cav3.2 and Cav3.3 were not detectable. (B) In Aqp4^–/– ANSCs, the mRNAs of genes encoding Cav1.2 and Cav3.1 were significantly decreased when normalized to GAPDH (^*P<0.05, n=3). There was no significant inhibition in the transcription of TRPC1 in Aqp4^–/– ANSCs compared with the Aqp4^+/+ control. (C) Western blot confirms that AQP4 knockout decreased the expression of Cav1.2 channel protein in the ANSCs compared with the wild type (^*P<0.05, n=3). Cav3.1 channel protein was not detected. (D) Western blotting (top panel) showing that the expression level of Cx43 protein in Aqp4^–/– ANSCs decreased to 39% of the level in the Aqp4^+/+ genotype (lower panel) (^*P<0.01, n=3).