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. 2021 Nov 8;9:741975. doi: 10.3389/fcell.2021.741975

FIGURE 6.

FIGURE 6

Differential expression analysis of junction epithelial mRNAs between WT and Ankrd31–/– epididymides by qRT-PCR. (A) Ocln, (B) ZO-1, (C) Cldn5, (D) CDH1, and (E) Cx43 expression levels were normalized using RP18S as a housekeeping gene and expressed as normalized fold expression (n.f.e.). All data are reported as mean value ± S.E.M; **p < 0.01. Western blot analysis of (F) actin, (G) tubulin, (H) OCLN, (I) ZO-1, (J) CLDN5, (K) CDH1, and (L) Cx43 proteins levels in WT and Ankrd31–/– epididymides. Signals were quantified by densitometry analysis and normalized to Ponceau Red (Pon.S). Data are expressed in OD values as fold change and reported as mean ± SEM, *p < 0.05; **p < 0.01. (M) IP in WT epididymides. Total proteins collected from WT epididymides were immunoprecipitated using ANKRD31 antibody. Protein interaction among ANKRD31, SPECTRIN, CDH1, ZO-1, and OCLN was detected by Western blot analysis.