Fig. 3
GM-CSF stimulates phagolysosomal fusion and IL-1β secretion in the absence of antimicrobial peptide production or NO production. a Nitrite production by uninfected, Mtb-infected, and GM-CSF stimulated Mtb-infected macrophages. Representative of three donors. b Quantitative PCR analysis of CAMP mRNA expression normalized to GAPDH expression in primary human macrophages were treated with GM-CSF or active Vitamin D. Each value represents the mean of ± standard deviation of three independent experiments. c Phagolysomal colocalization of GFP-Mtb with Lysotracker red was measured in 200–600 macrophages per well. Representative of three donors. d Small molecule GM-CSF inhibitor screen example. Bars pointing to the right identify drugs that had an increase in bacterial control while bars pointing to the left indicate a decrease in the setting of GM-CSF. e The effect of glibenclamide on GM-CSF mediated control was compared for six independent experimental donors (each donor independently color-coded). f The effect of glibenclamide or glipizide on GM-CSF mediated control was compared between vehicle-, glipizide- or glibenclamide-treated cultures. Representative of three donors. g Extracellular IL-1β production was measured from unstimulated and GM-CSF stimulated Mtb-infected macrophages. Error bars represent standard deviation of the technical replicates. Representative of three donors. h Extracellular IL-1β production was measured from unstimulated and GM-CSF stimulated Mtb-infected macrophages with small molecule inhibitors. Error bars represent standard deviation of the technical replicates. Representative of three donors. i LDH absorbance values in Mtb-infected macrophages with vehicle, GM-CSF or nigericin. Error bars represent standard deviation of the technical replicates. Representative of three donors. j Quantification of bacterial survival in the setting of GM-CSF with isotype or IL-1β antibody. Error bars represent standard deviation of the technical replicates. Representative of three donors. All comparisons made using a two-tailed t-test. (ns, not significant, **p < 0.01, ***p < 0.001, ****p < 0.0001). Source data are provided as a Source Data file
