Fig 2: CXCR7 plays a protective role in the brain-mediated Hippo signaling pathway. (A) The 10 most significantly enriched ischemia/reperfusion (I/R)-associated pathways with downregulated CXCR7 expression. (B) Representative protein expression levels of TCF7L2, p-YAP, and p-TAZ detected at 24 h following surgery in the sham, MCAO, and shRNA-CXCR7 + MCAO groups (n = 3). (C) Densitometric quantification of TCF7L2/ß-actin. (D) Densitometric quantification of p-YAP/ß-actin. (E) Densitometric quantification of p-TAZ/ß-actin. *p < 0.05 versus the sham group, # p < 0.05 versus the MCAO group

Fig 3: The CXCR7/miR-182 axis affects ischemic/reperfusion (I/R) progression mediated through TCF7L2. (A) All downregulated miRNA gene regulatory networks. (B) Putative miR-182 binding sequence in the 3' untranslated region (UTR) of TCF7L2 mRNA. A mutation was generated in the TCF7L2 3'UTR sequence at the complementary site of the seed region in miR-182. A TCF7L2 3'UTR fragment with a wild-type (TCF7L2-Wt) or a mutant (TCF7L2-Mut) miR-182-binding sequence was cloned downstream of the luciferase reporter gene. (C) The TCF7L2-Wt or TCF7L2-Mut reporter vector cotransfected into C6 cells with vehicle (control), miR-182 mimic, or negative control (miRNA-Scr). The normalized luciferase activity in the control group was set as the relative luciferase activity. The data are presented as the mean ± standard error of the mean (n = 3). (D) Representative western blot bands showing TCF7L2 expression in the miRNA-Scr, miR-182 mimic, anti-miRNA-Scr, and anti-miR-182 groups of C6 cells (n = 3). (E) Densitometric quantification of TCF7L2/ß-actin. (F) Representative western blot bands showing TCF7L2 expression in the shRNA-Scr, shRNA-CXCR7, shRNA-CXCR7 + miRNA-Scr, and shRNA-CXCR7 + miR-182 mimic groups of C6 cells. (G) Densitometric quantification of TCF7L2/ß-actin. *p < 0.05 versus the miRNA-Scr group