Fig. 4.

Myeloid UBXN6 is essential for enhancing ERAD and inhibiting inflammation induced by damaged mitochondria in LPS-primed macrophages. A, B Expression levels of Sel1l, Edem1, Syvn1, Dnajc10, and Herpud1 mRNAs in BMDMs measured by qRT‒PCR after treatment with LPS (100 ng/mL) (A) or SMER28 (20 μM) (B) for the indicated times. C Representative immunoblots showing the time course of SEL1L and SYVN1 protein induction in BMDMs. ACTB represents the loading control. D Relative quantitative analysis of SEL1 or SYVN1 levels normalized to ACTB in (C). E, F Representative images (E) and quantification (F) of mtROS levels in BMDMs stained with MitoSOX (1 μM) for 20 min after stimulation with LPS (100 ng/mL) for the indicated times. G The expression levels of Tnf and Il1b mRNAs in BMDMs after 1 h of pretreatment with or without MitoTEMPO (1 or 10 μM). The BMDMs were then stimulated with or without LPS (100 ng/mL) for 6 h. Statistical significance was determined via one-way ANOVA with Tukey’s multiple comparison test (A, B, F, and G) or two-tailed Student’s t test (D). SC, solvent control; LPS, lipopolysaccharide; n.s., not significant; a.u., arbitrary unit; MT, MitoTEMPO. The data represent the means ± SD (A, B, D, F, and G) from at least three independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001