Extended Data Fig. 5: FH deletion increases bioenergetic stress, fumarate, and mitochondrial membrane potential.
From: Macrophage fumarate hydratase restrains mtRNA-mediated interferon production
a, Bioenergetic ratios in BMDMs treated with DMSO or FHIN1 (n = 3). b, Fumarate and 2SC levels in BMDMs treated with DMSO or FHIN1 (n = 3). qPCR (n = 5) (c) and western blot (n = 2) (d) analysis of Fh1 expression in Fh1+/+ and Fh1–/– BMDMs (EtOH/TAM 72 h; LPS 4 h; Fh1+/+ NS vs Fh1+/+LPS (P = 0.00000002), Fh1+/+ NS vs Fh1–/– NS (P = 0.00000000000002), Fh1–/– NS vs Fh1–/– LPS (P = 0.0000000000014)). e, Bioenergetic ratios in Fh1+/+ and Fh1–/– BMDMs (n = 3; EtOH/TAM 48 h). f, Heatmap of top 50 significantly abundant metabolites in Fh1+/+ and Fh1–/– BMDMs (n = 3; LPS 4 h). g, Fumarate and 2SC levels in Fh1+/+ and Fh1–/– BMDMs (n = 3; EtOH/TAM 72 h). h, Glycolysis as measured by ECAR in BMDMs pre-treated with DMSO, FHIN1 or DMF (n = 8 (DMSO/FHIN1); (n = 6 (DMF); LPS 4 h). n = technical replicates from 1 experiment performed with 3 pooled biological replicates. Data are mean ± s.d. i, Glyceraldehyde 3- phosphate (G3P) and 2,3-phosphoglycerate (2/3-PG) levels and ratio in BMDMs pre-treated with DMSO or FHIN1 (n = 3; LPS 4 h; G3P (P = 0.00004)). Immunofluorescence (j) and quantification (k) of Mitotracker red staining in BMDMs pre-treated with DMSO or FHIN1 (n = 8 (DMSO); n = 19 (FHIN1); LPS 4 h). n = technical replicates from representative experiment. Scale bar = 20 μm. Data are mean ± s.d. a-c,e,g,i Data are mean ± s.e.m. Representative blots or images of 2 (d) or 1 experiment(s) (j) shown. n = biological replicates unless stated otherwise. P values calculated using two-tailed Student’s t-test for paired comparisons or one-way ANOVA for multiple comparisons.
