Fig. 4: RNA sequencing on non-treated, NTC and NRIP1KO mature adipocytes on day 6 after differentiation.
From: CRISPR-enhanced human adipocyte browning as cell therapy for metabolic disease
a Principal component plot prior to DEseq analysis. b Volcano plot of upregulated (red) and downregulated (blue) genes between control unedited adipocytes (NT and NTC) and NRIP1KO adipocytes with highlighted (green) Ucp1, Cidea, Fabp3, Nrg4. c Top 10 pathways associated with all upregulated genes detected. d Heatmap of pathway analysis of upregulated genes ranked according to the p value. e Heatmap of the 78 genes associated with cellular respiration that were upregulated in NRIP1KO adipocytes. f Heatmap of genes related to thermogenesis. Arrow shows Ucp1. g Browning probability calculated using the ProFAT online tool. p = 0.0004 by One-way ANOVA. h Left: top 15 upregulated genes by RNA sequencing where all NRIP1KO samples have >1000 normalized reads and padj > 0.1 (Black = NTC + NT) and NT, (Red = NRIP1KO). Right: Screening of these 15 genes in the excised implant NTC (black) or NRIP1KO (red) tissue by RT-PCR. P values *Ucp1 = 0.011; **Otop1 = 0.0017; *Cidea = 0.013; *Acaa2 = 0.018 by unpaired two-tailed T-test. Cut-offs were set at padj < 0.1 and fold change > 1.3. NT (n = 3), NTC (n = 5), NRIP1KO (n = 5). NRIP1KO adipocytes were transfected with SpyCas9 and either Nrip1 sgRNA-M4 or sgRNA-M6.
