Adipocytes undergoing ERS release functional molecular parts to induce the UPR in hepatocytes. (A) Schematic of the experimental design. (B) Agarose gel of Xbp1 cDNA amplicons from Ctrl/CM-treated AML12. Cells with no treatment (Blank) or 0.2 μM Tg treatment (Tg) are for controls. U, unspliced Xbp1; S, spliced Xbp1. (C) RT-qPCR of mRNA of the UPR markers in AML12 treated as in B. The statistical analyses were performed using an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001. (D) Upper panel, western blot of p-IRE1α, XBP1s, p-PERK, and BiP in AML12 treated as in B. p-PERK was determined by a slower band shift due to phosphorylation. The diamond indicates the nonspecific band. Lower panel, quantification of protein levels. Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were performed using an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. (E) KEGG enrichment analysis of genes with |log₂ (fold change)| >0.5 in AML12 treated with CM for 6 h, with Ctrl treatment as the control. FDR, false discovery rate. (F) Upper panel, western blot of p-IRE1α, XBP1s, p-PERK, and BiP in primary mouse hepatocytes treated as in B. p-PERK was determined by a slower band shift due to phosphorylation. The diamond indicates the nonspecific band. Lower panel, quantification of protein levels. Lanes were numbered from left to right, with the ratio in lane 2 (Tg treatment) set as 100. The statistical analyses were performed using an unpaired t test, two-tailed. Data were shown as the mean ± SEM, n = 3. *P < 0.05, **P < 0.01. Source data are available for this figure: SourceData F1.