Knocking down PRLR using Ad-shPRLR decreases insulin (Ins) sensitivity in vivo. Male C57BL/6J mice were infected with Ad-shPRLR
(+ Ad-shPRLR) or scrambled vector (− Ad-shPRLR) via tail-vein injection, followed by examination of PRLR expression in the
liver at day 7 in A, measurement of blood glucose and serum insulin levels at day 14 in B and C, calculating HOMA-IR index in D, performance of GTTs and ITTs at day 3 or 4 in E, and examination of insulin signaling in liver before (− Ins) and after (+ Ins) 2 units/kg insulin stimulation for 3 min
at day 7 in F. The mean ± SEM values shown are representative of at least two independent in vivo experiments, with the number of mice
included in each group in each experiments indicated (n = 5–6). Statistical significance was calculated using the two-tailed Student t test for the effects of the Ad-shPRLR vs. the control group (*P < 0.05). A: Prlr mRNA and PRLR protein (top, Western blot; bottom, quantitative measurement of PRLR protein relative to actin). B: Blood glucose levels. C: Serum insulin levels. D: HOMA-IR index. E: GTT and ITT. F: p-IR (tyr1150/1151), p-Akt (ser473), and PRLR protein (left, Western blot; right, quantitative measurements of p-IR and p-Akt protein relative to their total protein). t, total.