Fig. 7: Ubiquitination of K81 and K195 in SPSB2 are required for the regulatory effects of MDM2 on iNOS-NO axis and HIF-1α activation in M1 THP-1 macrophages. | Nature Communications

Fig. 7: Ubiquitination of K81 and K195 in SPSB2 are required for the regulatory effects of MDM2 on iNOS-NO axis and HIF-1α activation in M1 THP-1 macrophages.

From: MDM2 induces pro-inflammatory and glycolytic responses in M1 macrophages by integrating iNOS-nitric oxide and HIF-1α pathways in mice

Fig. 7

a S-KO-THP-1 and the Ctrl-THP-1 macrophages were transfected with the indicated plasmids, followed by immunoblotting analysis of SPSB2 and iNOS. n = 3. b–g S-KO-THP-1 and the Ctrl-THP-1 macrophages were co-transfected with siMDM2 or siScramble and plasmids encoding wild-type SPSB2 (WT) or the K81R or K195R mutants of SPSB2, followed by stimulation with LPS and IFNγ for 24 h. n = 3. b Total nitrate & nitrite in the conditioned medium. n = 4. c HIF-1α transcriptional activity in the nuclear fraction of the cells. n = 4. d IL-1β and e lactate in the conditioned medium. n = 4. f, g QPCR analysis of pro-inflammatory genes (f) and glycolytic genes (g) normalized with β-ACTIN. The QPCR results were displayed as fold change over Ctrl-THP-1-siScramble-GFP. n = 4. Data are displayed as mean ± SEM. Statistical significance was examined using one-way ANOVA with Tukey post-hoc test. N number represents the number of biological replicates. Representative immunoblot images are shown.

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