PRLR regulates insulin (Ins) sensitivity in vitro. A and B: Cells were infected with Ad-PRLR (+ Ad-PRLR) or GFP (− Ad-PRLR) for 48 h in A. HepG2 cells were treated with PRLR siRNA (+ PRLR RNAi) or control reagent (− PRLR RNAi) for 48 h, and primary hepatocytes
were infected with Ad-shPRLR (+ Ad-shPRLR) or scrambled vector (− Ad-shPRLR) for 72 h in B; both cases followed with (+ Ins) or without (− Ins) 100 nmol/L insulin stimulation for 20 min. C and D: Primary hepatocytes were exposed to Ad-PRLR (+ Ad-PRLR) or were without Ad-PRLR (− Ad-PRLR) for 24 h. The cells were then
treated with or without 100 nmol/L insulin for another 24 h, followed by the measurement of glucose production or glycogen
content. The mean ± SEM values shown are representative of at least three independent in vitro experiments. Statistical significance
was calculated using one-way ANOVA followed by the SNK test for the effects of any group vs. the − Ad-PRLR group without insulin
stimulation (*P < 0.05), with vs. without insulin stimulation in the + Ad-PRLR group (#P < 0.05), or Ad-PRLR vs. the control group after insulin stimulation (&P < 0.05). A and B: p-IR (tyr1150/1151), p-Akt (ser473), and PRLR protein (top, Western blot; bottom, quantitative measurements of p-IR, p-Akt, and PRLR protein relative to their total protein or actin). C: Glucose output assay. D: Glycogen content. t, total.