Fig. 3.

LncRNA-Fendrr may regulate the ubiquitination of NLRC4 protein through HERC2. a The expression of Fendrr was detected by qRT-PCR. b, c After transfection, the cells were treated with Cycloheximide (CHX) (50 μg/mL) for 0, 2, 4 and 6 h, and the NLRC4 protein level was detected through western blot. d, e The binding of lncRNA Fendrr to HERC2 protein was detected by (d) RNA pull-down and (e) RNA Immunoprecipitation (***P < 0.001 versus the IgG group). f The interaction between HERC2 and NLRC4 was detected by Co-Immunoprecipitation. g, h pcDNA3.1-HERC2 or pcDNA3.1 (control) was transfected into BV-2 cells, and then HERC2 and NLRC4 were detected by Western blotting. i, j si-HERC2 or si-NC (control) was transfected into BV-2 cells, and then HERC2 and NLRC4 expressions were detected by Western blotting. k BV-2 cells were treated with MG132 (10 μg/mL) and transfected or co-transfected with pcDNA3.1-Fendrr and pcDNA3.1-HERC2 to detect NLRC4 ubiquitination levels. n = 3/group. Data are presented as means ± SD. P values were analyzed by Student’s t-test. **P < 0.01 and ***P < 0.001 versus the pcDNA3.1 group