Extended Data Fig. 2: Loss of Klf9 function impairs the cardiac mitochondrial function.
From: Klf9 is essential for cardiac mitochondrial homeostasis
a, Volcano plot indicating the genes that were differentially expressed in WT versus Klf9 KO hearts. In red are the genes significantly up-regulated and in blue are the genes significantly down-regulated. b, KEGG pathway enrichment analysis of downregulated genes in Klf9 KO hearts. Orange dots represent the number of genes of each annotated pathway and the length of each column is based on the p values. c, Volcano plots showing the GSEA of GO cellular component categories of differentially expressed genes (DEGs). Gene sets related to mitochondria are marked. Sizes of the dots represent the number of genes in the gene sets and the transparency represent the FDR of the enrichment. d, Top hits of GSEA analysis from c showing the Klf9 positive correlation with the mitochondria. NES, normalized enrichment score; FDR, false discovery rate. e, GSEA analysis of disease category of DEGs. f-i, Heat-map, qRT-PCR (n = 6) and Western blotting analysis (n = 3) of the expression of genes involved in FA oxidation, oxidative phosphorylation, mitochondrial biogenesis, mitochondrial dynamics, autophagy and mitophagy. j, ChIP assay of heart extracts showing the enrichment of Klf9 proteins at the promoter region of PGC-1α (n = 4). k, TEM analysis of hearts from the indicated mice. l, Quantitative data of mitochondrial size distribution and content based on TEM images (n = 6). m, n, Left, Flow cytometric analyses of size (forward scatter, FSC) (m) and transmembrane electrical potential (DiOC6 intensity) (n) of isolated cardiac mitochondria from the indicated hearts. Right, Quantitative data of FSC and DiOC6 intensity (n = 6). o, Left, Oxygen consumption rates (OCR) showing the respiration of isolated mitochondria from the indicated hearts. Right, Quantification of basal and FCCP induced maximal respiration (n = 9). p, Myocardial ATP levels in the indicated hearts (n = 12). q, Left, Mitochondrial ROS production of isolated mitochondria from the indicated hearts measured with MitoSOX. Right, Quantitative data of ROS intensity (n = 6). Data are shown as mean ± SEM; 2-tailed Student’s t test (g, h, i, l, m, n, p, q), 2-way ANOVA (j, o).
