Fig. 1: TRIM22 actives NF-κB signaling via K48-linked ubiquitination of IκBα.
From: TRIM22 activates NF-κB signaling in glioblastoma by accelerating the degradation of IκBα
a Luciferase activity from U87MG and LN229 cells transfected with siRNAs of TRIM5, TRIM21, TRIM22, TRIM38, or TRIM56, along with a reporter plasmid carrying the NF-κB promoter relative to negative control. b Western blot analysis to evaluate core kinases in the NF-κB pathway in lysates (20 µg) prepared from TRIM22 knockout cells. GAPDH was used as the loading control. c qRT-PCR analysis of IκBα mRNA levels in modified U87MG and LN229 cells relative to the control sg-scramble. GAPDH was used to normalize samples. d Western blot analysis of IκBα protein in TRIM22-depleted cells treated with cycloheximide (CHX; 25 μg/mL) for 0, 8, 16, and 24 h. e Decay curve of IκBα levels normalized to β-tubulin and to 0 h at the indicated time points from CHX experiments. f Western blot analysis of IPs performed with antibody to IκBα to detect endogenous IκBα ubiquitination from indicated cells. Antibody for K48-linked polyubiquitin was used on the western blot. Student’s t test: n.s. not significant, *P < 0.05.
