Fig. 5

Interaction with RNA binding proteins induces LncRNA packaging into adipocyte exosomes. a. Quantitative real-time PCR analysis shows the relative level of cellular and exosomal LOC606724 (LOC) and SNHG1 in nADs or MMADs. b. Western blotting shows the level of hnRNPA2B1 or hnRNPU in the lysate of MMADs using RNA pull down by biotin-labeled sense or antisense transcript of LOC. Inputs served as control. c & d. RIP assay shows the relative enrichment of LOC or SNHG1 in the immunoprecipitates of MMAD lysates pulled down by anti-hnRNPA2B1 (c) or anti-nRNPU (d) antibodies. e. RNA interference efficacy of sihnRNPA2B1 and sihnRNPU in MMADs. f. Quantitative real-time PCR analysis shows the relative level of exosomal LOC and SNHG1 derived from MMADs carrying sihnRNPA2B1 or sihnRNPU. g. Annexin V-binding assay shows the percentages of apoptotic ARP-1 cells treated with 5 nM bortezomib (BTZ) and exosomes derived from MMADs carrying sihnRNPA2B1 or sihnRNPU. ARP-1 cells treated with vehicle or exosomes derived from MMADs carrying siCtrl served as controls. h & i. RIP assay shows the relative enrichment of LOC or SNHG1 in the immunoprecipitates from lysates of nADs or MMADs pulled down by anti-hnRNPA2B1 (h) or anti-hnRNPU (i) antibodies. Cell lysates pulled by IgG served as control. Data shown as mean ± SD. ns, non-significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. P values were determined by Student t-tests for comparison of two groups and one-way ANOVA for comparison of more than two groups