FIG. 6.

Analysis of the CF content and its action mechanism. A and B: CFs, inactivated by proteinase-k (CFp) or by RNase cocktail (CFr), were delivered into oASC. Glucose uptake and the cell size-to-cell number ratio were measured in mASCs (n = 5). *P < 0.05. C: Analysis of Lin28 protein expression by Western blot in oASC-derived or cASC-derived CFs or in whole ASC lysates to note the restoration of Lin28 levels after cASC-derived CF (cCF) delivery into oASC (mASC). One representative image of three independent experiments is shown. D: Let7 microRNA expression was analyzed by RT-PCR (n = 5). *P < 0.04. E: Insulin-stimulated glucose uptake by adipocytes differentiated (7 days) from modified oASCs or oASC delivered with Lin28 protein. Adipocyte cultures were maintained overnight in serum-free, low-glucose medium and then stimulated with 10 nmol/L insulin (30 min; n = 5). *P < 0.03 between oASCs and mASCs or oASCs+Lin28. F: Representative images of Oil Red O–stained oASCs and oASC delivered with Lin28 protein after 7 days of differentiation into mature adipocytes (n = 5). Bar, 30 μm. G: Adiponectin, MCP-1, and TNF-α production by oASC-derived adipocytes (7 days) transferred or not with Lin28 and detected by immunoassay. Results are expressed as the percentage of detection comparing undifferentiated oASCs at day 0 to differentiated oASCs at day 7 (n = 5). *P < 0.02. H: Let7 microRNA and Lin28 expression were analyzed by RT-PCR in cASC-derived adipocytes (7 days) in the presence or absence of 1 ng/mL TNF-α or 0.2 ng/mL MCP-1 cytokines and in oASC-derived adipocytes in the presence or absence of 10 ng/mL anti–TNF-α or anti–MCP-1 antibodies (Abs) (n = 5). *P < 0.05 and **P < 0.02. d, days; Fibrob, fibroblast.