Fig. 6

Nudt21 Impedes Autophagy’s Role in Reducing Inflammatory Cytokine Production. A, B Histograms displaying cytokine levels in BMDMs from WT and Nudt21-cKO mice primed with 50 ng/ml IFNγ overnight and 50 ng/ml LPS for 8 hours (n = 3 per group): (A) IL6 gMFI. (B) TNFα gMFI. ELISA results showing the IL6 (C) and TNFα (D) levels in the supernatants of the cultures primed with 50 ng/ml IFNγ overnight and 50 ng/ml LPS for 12 hours, with or without 100 nM BafA1. (n = 5 per group). RT‒qPCR analysis of cytokine mRNA levels in BMDMs primed with 50 ng/ml IFNγ overnight and 50 ng/ml LPS for 8 hours: (E) IL6 mRNA levels (n = 3 per group). (F) TNFα mRNA levels (n = 3 per group). G Immunoblot analysis of phosphorylated IKKα/β, total IKKα, total IKKβ, phosphorylated NF-κB p65 (p-p65), total p65, p62, LC3, and NUDT21 levels in WT and Nudt21-cKO BMDMs primed with 50 ng/ml IFNγ overnight and 50 ng/ml LPS, with or without 100 nM BafA1, for the indicated durations. β-ACTIN was used as the loading control. The error bars represent the standard errors of the means (SEMs). P values were determined by unpaired Student’s t-test