Fig. 6

Silencing RCN3 induces ER stress in MMCs through calreticulin (CALR) and mesencephalic astrocyte derived neurotrophic factor (MANF) suppression. A, B MMCs were transfected with RCN3 siRNA (20 nM) for 24 h, and then treated under NG condition for 48 h. CALR and MANF mRNA and protein expression were measured. C, D MMCs were treated with HG-treated HK-2 exosomes for 48 h. CALR and MANF mRNA and protein expressions were examined. E, F ATF-6, CHOP, phospho-PERK and phospho-IRE1α protein expressions were assessed in MMCs transfected with CALR or MANF siRNA for 24 h, and then treated under NG condition for 48 h. G, H RCN3 mRNA and protein expression were assessed in MMCs transfected with CALR or MANF siRNA for 24 h, and then treated under NG condition for 48 h. MANF mRNA and protein expression were measured in MMCs transfected with CALR siRNA for 24 h, and then treated under NG condition for 48 h. CALR mRNA and protein expression were measured in MMCs transfected with MANF siRNA for 24 h, and then treated under NG condition for 48 h. mRNA and protein level were measured using qPCR and Western blotting respectively. The bar graph represents the mean ± S.E.M. of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 by Student’s t-test