Skip to main content
. Author manuscript; available in PMC: 2018 Sep 1.
Published in final edited form as: J Immunol. 2017 Jul 28;199(5):1805–1816. doi: 10.4049/jimmunol.1700515

Figure 2. iNOS is required for HIF-1α stabilization and transcriptional activity.

Figure 2

(A) RNAseq reads for Hif1a 24h post-infection in wildtype and Nos2 BMDM in untreated [un], M. tuberculosis infected [TB], and M. tuberculosis infected with IFN-γ activation [TB/G]. (B) Western blot for HIF-1α in M. tuberculosis infected, IFN-γ activated BMDM at 0,4,12 and 18h post-infection in wildtype and Nos2−/− BMDM. Values for quantification of HIF-1α/tubulin ratios are normalized to lane 3 (maximal HIF-1α levels) and are shown below the image. (C) Western blot for HIF-1α during infection with M. tuberculosis with IFN-γ activation in the presence or absence of ascorbate (D) Western blot for HIF-1α in M. tuberculosis infected, IFN-γ activated BMDM with a dose response of the iNOS inhibitor 1400W [.625, 1.25, 2.5, 5, 10, 25uM]. Griess assay for NO production was done on the supernatant of the cells used for the western blot. (E) Western blot for HIF-1α in M. tuberculosis infected, IFN-γ activated BMDM with a dose response of 1400W [.625, 1.25, 2.5, 5uM] with and with addition of the NO donor SNAP [250uM]. (F) HIF-1α levels were measured by western blot at 12h after stimulation with PAM [50ng/ml] and IFN-γ [6.25 ng/ml]. RNAseq data is from 3 independent experiments. All other experiments are representative of 2 or more experiments. Error bars represent the SD and p values were determined using an unpaired t test. ***p<.001