Journal: Molecular metabolism

Article Title: The transcriptional co-regulator LDB1 is required for brown adipose function.

doi: 10.1016/j.molmet.2021.101284

Figure Lengend Snippet: Figure 4: Glucose tolerance and uptake are reduced upon loss of LDB1, in part via altered insulin signaling. (A) Plasma blood glucose from male (n ¼ 5) and female (n ¼ 11) CTL and Ldb1DBAT mice after a 6-h fast. (B) Intraperitoneal glucose tolerance tests (IPGTT) after injection of 2.0 g/kg body weight of glucose for CTL and Ldb1DBAT mice raised at 28 C for female mice (CTL n ¼ 11 and Ldb1DBAT n ¼ 12) and male mice (CTL n ¼ 5 and Ldb1DBAT n ¼ 4) aged 2e4 months. Inset shows Area Under the Curve (AUC). (C) Intraperitoneal insulin tolerance tests (IPITT) to assess changes in blood glucose upon injection of 0.5U/kg body weight of insulin in CTL and Ldb1DBAT male mice at 28 C (n ¼ 5e6). (DeE) IPITT conducted in male mice 15 min after an acute 4 C cold challenge or CL316,243 injection, respectively (n ¼ 5e6; aged 3-month-old). Genotype effect found on lower right corner calculated via 2-way ANOVA. (F) Glucose uptake via 2DG glucose tracer in male mice (n ¼ 5e7; 8e9-month-old). (G) Plasma insulin assessed via ELISA from CTL and Ldb1DBAT male mice at 28 C (n ¼ 6). (H) Assessment via western for pan or pAKTS473 protein (n ¼ 3); Bottom panel is densitometry of pAKTS473 protein band intensity normalized to pan AKT. (I) Assessment via western of total and p-Erk1/2 protein (n ¼ 3); Bottom panel is densitometry of p-Erk1/2 protein band intensity normalized to total Erk1/2 protein. *, p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant via Student’s t-tests or two-way ANOVA, where appropriate.

Article Snippet: Polyvinylidene difluoride (PVDF) membranes were probed using the following primary antibodies: rabbit a-LDB1 (1:1000, generously provided by Dr. Paul Love, NIH), rabbit a-UCP1 (1:1000, Cell Signaling #14670S (D9D6X)), mouse a-Actin (1:1000, Abcam ab3280), rabbit apAKTS473 (1:1000, Cell Signaling #9271S), rabbit Pan AKT (1:3000, Cell Signaling #4691S), rabbit a-phospho-p44/42 MAPK(T202/4204) (1:1000, Cell Signaling #4370S), and rabbit a-p44/42 MAPK(T202/4204) (1:1000, Cell Signaling #4695S).

Techniques: Clinical Proteomics, Injection, Enzyme-linked Immunosorbent Assay, Western Blot