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. 2022 Mar 28;2(4):100192. doi: 10.1016/j.crmeth.2022.100192

Figure 4.

Figure 4

Metabolic analysis of BMDMs and HMDMs with SCENITH reveals expected macrophage activation by LPS ± IFNγ, and IL-4

(A and B) MFI of puromycin across samples treated with different inhibitors for BMDMs (A) and HMDMs (B). DG, O and DGO indicate Deoxyglucose- (DG), oligomycin- (O), or deoxyglucose + oligomycin-treated (DGO) samples.

(C) Calculations of metabolic SCENITH parameters based on puromycin MFI.

(D, E, F, and H) SCENITH parameters as calculated for mouse (D and F) and human (E and H) macrophages.

(G and I) Correlation of glycolytic capacity as measured with XF analysis with glycolytic capacity as measured with SCENITH for BMDMs (G) and HMDMs (I).

(J and K) tSNE dimensionality reduction of naive, LPS ± IFNγ-, and IL-4-treated BMDMs (J) and HMDMs (K) and clustered heatmaps showing the expression of activation markers and puromycin per stimulus.

Data are shown as mean ± SEM Each dot marks a separate mouse (n = 6) or human donor (n = 6). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by two-way (A and B) or ordinary one-way ANOVA(D, E, F, and H) with Dunnett’s post hoc test for multiple comparisons. Correlations were fitted using a simple linear regression model (G and I).