Fig. 7: Hypoxia in kidneys of Il1r1-/- mice promotes pathogenicity of C. albicans. | Nature Communications

Fig. 7: Hypoxia in kidneys of Il1r1-/- mice promotes pathogenicity of C. albicans.

From: IL-1 protects from fatal systemic candidiasis in mice by inhibiting oxidative phosphorylation and hypoxia

Fig. 7

A ECAR and OCR B values from a Seahorse XF96 ATP assay analysis of differentiated AB8/13 cells, treated overnight with either IL-1b, Anakinra or neither. 5 well per condition were analyzed (representative of two independent repeats is shown). Data are presented as mean values with +/- SD. C Calculated mitochondrial/glycolytic ATP source from the experiment in (A, B). Calculations were performed as described in the Seahorse XF user guide47 (5 well per condition were analyzed, representative of two independent repeats is shown). Statistical test: two tailed unpaired t tests on MitoATP/GlycoATP, comparing stimulation to control. P[Il-1b] = 0.0005, P[Anakinra] < 0.0001. Data are presented as mean values with +/- SD. D Kidney sections from infected Il1r1+/- (top) and Il1r1-/- (bottom) mice, 24 h after infection with 105 CFU C. albicans stained with antibodies against Pimonidazole (green) and C. albicans (red) and with Hoechst (blue). Merged images of 2 color-channels (Hoechst and Cy5) are shown. While both were stained using the same secondary antibody (fluorophore Cy5), different colors are chosen to display Pimonidazole and C. albicans staining. E Quantification of the high hypoxia areas from (D). Total area of high hypoxia was calculated using ImageJ2 (version 2.14.0) and normalized to the total area of the kidney section. To define high hypoxia, a color threshold was defined for green (Pimonidazole), with brightness above physiological hypoxia. All samples were analyzed using the same color and brightness thresholds. Each point represents an animal, n = 4. Statistical test: two tailed unpaired t test on normalized areas. P = 0.0237. Data are presented as mean values with +/- SD. F C. albicans colonies grown on YPD agar plates for 48 h, 37 °C, 5% CO2, in normal atmospheric oxygen levels (left) and in hypoxia (1% O2) (right). Imaging done by light microscopy. (A representative picture from a single experiment with n = 3 plates per condition is shown).

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