Figure 4.

VitA mediates the pro-fibrotic transcriptional response in kidney tissue following HFD feeding independent of mitochondrial energetics. Data are reported as mean values ± SE. Two-way ANOVA was performed to analyze differences by HFD feeding and VitA. Results of post-hoc analyses for each comparison are summarized by symbols as defined: # p<0.05 for HFD feeding, $ p<0.05 for VitA, and & p<0.05 for the interaction between HFD feeding and VitA. V_ADP of isolated kidney mitochondria with (A) palmitoyl-carnitine (#) and (B) pyruvate each combined with malate as substrates (n=6-8). (C) Citrate synthase and (D) HADH activity in kidney tissue (n=7-8). (E) Representative immunoblots and (F) densitometric analysis of NDUFA, SDHA (&), ATP5A (&), MnSOD, UCP3 (#, $, &), and 4-HNE normalized to Vinculin. Data are presented as fold change relative to NCD (assigned as 1.0; dashed line), n=6. (G) mRNA expression of transcripts involved in mitochondrial energetics (Cs: #; Cpt1b: $; Hadha: #; Hadhb: $, &). Data are presented as fold change relative to NCD and normalized to Rps16 (assigned as 1.0; dashed line), n=8. (H) Representative H&E stains of kidney sections (scale bar: 50 μm). (I) mRNA expression of transcripts involved in organ fibrosis (Col1a1: &; Col3a1: &). Data are presented as fold change relative to NCD and normalized to Rps16 (assigned as 1.0; dashed line), n=8. *p < 0.05 vs. normocaloric diet same VitA availability, † p<0.05 vs. VitA sufficiency same caloric diet.