Fig. 7: Conserved and cold-inducible PPARGC1A-AT drives Adcy3-at expression in BAs.
From: Cold-induced expression of a truncated adenylyl cyclase 3 acts as rheostat to brown fat function
a, Sashimi plots visualizing splicing junctions from aligned RNA-seq data in BAT in Ppargc1 from 20-week-old male C57BL/6N mice at 22 °C or after 24 h of 5 °C CE; n = 3–5. Illu, Illumina short-read RNA-seq; Telo, TeloPrime full-length cDNA-seq; cDNA, direct cDNA-seq. Reads were aligned against GENCODE M29 annotation and transcript reassembly using Illumina short-read and full-length RNA-seq using FLAIR93. b, Schematic of the canonical Ppargc1a transcript and PPARGC1A protein structure (1) and Ppargc1a-at mRNA and PPARGC1A-AT protein structure (2). Created with BioRender.com. c, Genomic conservation of Ppargc1a-at among representative mammals. Colour gradient indicates the percentage of nucleotide identity of exon 1b relative to mouse. Phylogeny based on ref. 89. d–f, Relative expression of Ppargc1a (blue) and Ppargc1a-at (red) as determined by qPCR analysis of primary adipocytes derived from SVF cells from BAT (d), iWAT (e) and eWAT (f) depots. Replicates represent primary adipocytes isolated from individual mice (n = 3). Bar graphs represent the mean + s.e.m. with all data points plotted. To test for statistical significance, non-parametric (ranked) Kruskal–Wallis one-way ANOVA tests with Dunn’s correction for multiple testing were performed. P values are indicated. g, Expression of human PPARGC1A-AT in human 1°BAs derived from SVF precursors and stimulated with 1 µM NE for 16 h (n = 6 per condition), as described previously89. An unpaired, two-tailed and non-parametric Mann–Whitney test was performed to assess statistical significance. Data are shown as a percentage as the mean + s.e.m. with all data points plotted. The P value is indicated. h–k, Expression of Adcy3-at (h), Adcy3 (i), Ppargc1a-at (j) and Pparg1a (k) in mouse 1°BAs after transfection with 25 nM scrambled (scr) LNA inhibitors targeting both Ppargc1a (LNA_2) isoforms or exclusively Ppargc1a-at (LNA_1) and stimulated with 10 µM CL316,243 for 6 h. Data represent three to four independent experiments, each performed in three technical replicates. Paired samples are represented by individual lines and scr LNA set to unity. Line graphs represent the mean + s.e.m. with all data points plotted. Paired, two-tailed Student’s t-tests were performed to assess statistical significance. P values are indicated.
