Extended Data Fig. 2. Organelle mass and mitochondrial morphology are selectively altered in inflammatory macrophages.
a Quantification of organellar volume upon LPS/IFN-γ treatment. Data represent N = 59 (0 h), N = 48 (1 h), N = 56 (6 h), N = 46 (16 h), N = 59 (24 h) cells from n = 3 (16 h, LD 0–6 h) and n = 4 (non-LD 0–6, 24 h) independent experiments. P values were calculated using one-way ANOVA with Dunnett’s post-hoc test. b Cell size of naïve and 24h LPS/IFN-γ-treated BMDMs representing n = 6 independent experiments. P value was obtained by two-tailed, unpaired t-test. c-e Images (c) and quantification (d, average mitochondrial volume/cell; e, mitochondrial sphericity) showing dynamics in mitochondrial morphology of LPS/IFN-γ-activated BMDMs. Images are single z-planes and representative of n = 3 independent experiments. Scale bars: 5 µm, 1 µm (magnification) (c). Data represent (d) N = 51 (0 h), N = 76 (1 h), N = 86 (6 h) and N = 80 (24 h) and (e) N = 55 (0 h), N = 78 (1 h), N = 71 (6 h), N = 79 (24 h) cells from n = 3 independent experiments. P values were calculated using one-way ANOVA with Tukey’s post-hoc test. f-g Western blot analysis (f) and quantification (g) showing protein levels and activation state of DRP1 upon LPS/IFN-γ treatment. Data represent n = 3 biological replicates. P values were calculated using one-way ANOVA with Sidak’s post-hoc test (g). h,i Images (h) and quantification (i) showing the localization of DRP1 to mitochondria (HSP60) in LPS/IFN-γ-activated BMDMs. Scale bars: 5 µm, 1 µm (magnification) (h). Data represent N = 26 (0 h), N = 32 (1 h), N = 30 (6 h), N = 30 (24 h) cells from n = 3 biological replicates. P values were calculated using one-way ANOVA with Tukey’s post-hoc test (i). j-m WB analysis (j,l) and quantification (k,m) showing protein levels of MFN2 (j-k) and OPA1 (l-m) upon LPS/IFN-γ treatment. Data represent n = 3 biological replicates. P values were calculated using one-way ANOVA with Sidak’s post-hoc test (k,m). Boxes represent 25th to 75th percentiles, whiskers 10th and 90th percentiles. Dots are outliers, median is shown as line (a,d,e,i). The bars show the mean ± SEM (b) or the mean ± SD (g,k,m). Numerical P values are indicated in Supplementary Information Table 4 (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, not significant (ns) P > 0.05). Source numerical data and unprocessed blots are available in source data.