Fig. 2

Conditioned medium (CM) derived from MIIP-downregulated CRC cells induces increased browning, lipolysis, and cytokine secretion in adipocytes. a The flowchart outlines the preparation of CM from HCT116 WT or MIIP^+/− cells and the subsequent treatment of differentiated mature adipocytes. b RT-qPCR analysis reveals the expression of browning-related genes in mature adipocytes after 24 h of treatment with CM from HCT116 (MIIP^+/− or WT) cells (Adipo: mature adipocytes; 6 biological replicates per group). c Immunoblots show the levels of mitochondrial proteins (UCP1, PGC-1α), a key enzyme in lipolysis (total/phospho-HSL), and a lipid droplet protective protein (PLIN1) in mature adipocytes treated with CM from HCT116 (MIIP^+/− or WT) cells for 24 h (H89-2HCl: 20 μM for 3 h; representative of 3 biological replicates per group). d Representative images of optical microscope and Oil Red O staining in mature adipocytes treated with CM from HCT116 (MIIP^+/− or WT) cells for 24 h (40 × for images with a 50 μm scale bar; 3 biological replicates per group). e Mature adipocytes were treated with the indicated CM for 24 h, then washed, and the medium was changed. After another 24 h, the concentrations of free fatty acids (Left, 8 biological replicates per group) and glycerol (Right, 6 biological replicates per group) in supernatants were detected (H89-2HCl: 20 μM for 3 h). f RT-qPCR analysis shows the expression of white adipocyte-related genes in mature adipocytes after the treatment described in e (6 biological replicates per group). g Concentrations of TNFα (Left, 6 biological replicates per group) and IL-6 (Right, 5 biological replicates per group) were measured in supernatants of mature adipocytes described in e. h RT-qPCR analysis shows the expression of TNFα and IL-6 mRNA in mature adipocytes after the treatment as described in e (6 biological replicates per group). All data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001