Fig. 4
Taok3 deletion or inhibition increases differentiation of macrophages and their production of pro-inflammatory response factors. a Schematic representation of the method employed to differentiate and polarize bone marrow derive- macrophages (BMDMs). Representative flow cytometry plot showing the purity of BMDM cultures, as assessed by the frequency of CD11 + , F4/80 + macrophages. b Heatmaps representing the LOG10 (Fold change) relative to Taok3.+/+ unpolarized M0 using the mean fluorescence intensity by flow cytometry. Statistical comparisons are made between genotypes between the same polarization regimens. c Quantification of the TNFα levels in BDMSs from b) using ELISA. d Gating strategy used to discern between GM-SCF grown BMDCs (CD11c +) and M-CSF differentiated BMDMs (F4/80 +). e Flow cytometric analysis of the mean expression (mean fluorescence intensity) of surface molecules and transcription factors in BMDCs and BMDMs. f Bar plot of the frequency of differentiated macrophages, depending on the phase at which the cells were treated with the inhibitor. Differentiation (Day 0–6) → Polarization (Day 6–7). g Heatmaps representing the LOG10 (Fold change) relative to DMSO → DMSO unpolarized M0 using the mean fluorescence intensity by flow cytometry. Statistical comparisons are made between genotypes between the same polarization regimens against the respective DMSO → DMSO group. h Quantification of the TNFα levels in BDMSs from g) using ELISA. Data representative of at least three independent experiments. Chart error bars represent mean ± SEM. Statistical analysis: a-h Multiple unpaired, bilateral T-tests. ns, non-significant. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001