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. 2014 Aug 12;9(8):e96211. doi: 10.1371/journal.pone.0096211

Figure 1. CypA deficiency reduces activated NF-κB/p65 (RelA) expression.

Figure 1

(A) Western blot analysis of CypA, NF-κB subunits and their inhibitory protein, IκBα. Cytoplasmic extracts isolated from MEFs and C2C12 cells were immunoblotted for IκBα, and GDPH was used as internal control. Nuclear extracts prepared from MEFs and C2C12 cells were subjected to Western blot analysis to detect p65/RelA, p50, RelB, and p52 expression levels. p84 served as an internal control. Ppia +/+ and Ppia −/− represent CypA WT and CypA KO, respectively. SC is a scrambled clone, and 1C2 and 2B6 are C2C12 CypA Kd clones. (B) Semi-quantitative analysis of relative expression levels of p65/RelA in MEFs (Ppia +/+ vs Ppia −/−) and C2C12 cells (WT, SC vs 1C2 and 2B6). (C) qRT-PCR analysis of relative mRNA expression levels of p65/RelA and p50 between Ppia +/+ and Ppia −/− MEFs. (D) p65/RelA expression levels in MEF cytoplasmic extract (CE) and nuclear extracts (NE) upon treatment cells with CsA and Sfa.