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. 2022 Jul 19;298(9):102283. doi: 10.1016/j.jbc.2022.102283

Figure 1.

Figure 1

XBP1 modulates glucose and lipid metabolism in the liver. A, three sets of adenoviral shRNAs of XBP1 were added to Hepa1-6 cells for 48 h (n = 3). The bottom panel depicts the positions (blue square) of shRNAs’ targeting regions in XBP1. B, forty-eight hours after the addition of adenoviral shRNAs for scrambled control and XBP1 (ad-shXBP1-1), primary hepatocytes were subjected to serum starvation for 3 h, followed by glucose production medium supplemented with 0.2 mM cAMP for another 3 h (n = 3). C, adenoviral shRNAs for scrambled control and XBP1 (ad-shXBP1-1) were added 16 h after the seeding of Hepa1-6 cells. After 6-h incubation, 20 ng of reporter construct was transfected into Hepa1-6 cells together with 400 ng of control RSV-cat and PKA expression plasmids. Reporter activities were measured 48 h after the transfection (n = 3). D and E, three-month-old C57BL6 mice were injected with AAV8-scrambled shRNA or AAV8-XBP1shRNA (shXBP1-3) (3 × 10ˆ11GC/mouse). Sixteen days after the viral injection, blood glucose levels were examined after 8 h of fasting (D) (n = 8). Liver tissues were collected after 12 h of fast (E) (n = 5). F and G, three-month-old C57BL6 mice were injected with AAV8-scrambled shRNA or AAV8-XBP1shRNA through the jugular vein as in (D). Liver tissues were collected at 16 days after the viral injection, and triglyceride contents (F) (n = 7 ∼ 8) and the mRNA levels of genes related to lipid metabolism (G) were determined (n = 5). ∗p < 0.05, Student’s t test. AAV, adeno-associated viruses; SCD1, stearoyl coenzyme desaturase 1; XBP1, X-box binding protein 1.