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. Author manuscript; available in PMC: 2021 Feb 28.
Published in final edited form as: Mucosal Immunol. 2020 Aug 29;14(1):253–266. doi: 10.1038/s41385-020-00342-x

Figure 6: The effects of HO-1 inhibition on IFNγ-mediated control of intracellular bacterial levels correlate with intracellular iron levels.

Figure 6:

(A) CFU loads obtained from C57BL/6 BMDM 96 hours post infection and stimulation with IFNγ (250 U/ml), IFNγ (250 U/mL) plus SnPP (1μM), IFNγ (250 U/mL) plus SnPP (1μM) plus CORM2 (5μM) – IFNγ + SnPP + CO, IFNγ (250 U/mL) plus SnPP (1μM) plus biliverdin (5 μM) – IFNγ + SnPP + BV or IFNγ (250 U/mL) plus SnPP (1μM) plus FeSO4 (5 μM) – IFNγ + SnPP + Fe (triplicates); (B) NOS2 expression in C57BL/6 BMDM 24 hours post-infection and stimulation with IFNγ (100 U/mL), IFNγ (100 U/mL) plus SnPP (1μM) or IFNγ (100 U/mL) plus SnPP (1μM) plus FeSO4 (5 μM) – IFNγ + SnPP + Fe (triplicates); (C) Representative dot plots of data shown in Fig. 5B (flow cytometry data concatenated from triplicates); (D) Nitrite levels in supernatants of C57BL/6 BMDM infected and stimulated as in Fig. 5B (triplicates); (E) Labile iron pool readout measured as ΔRFU (relative fluorescence units) in BMDM lysates 24 hours post infection and stimulation or not with IFNγ (250 U/mL), SnPP (1μM) or IFNγ (250 U/mL) plus SnPP (1μM) (triplicates). The data are presented as the means ± standard error (A, B, D and E) or histograms from concatenated data (C). Data shown are representative of 2 (A, B, C, D and E) independent experiments. Statistical analysis: Student’s t test. * = p<0.05, ** = p<0.01, *** = p<0.001, n.s. = non-significant.