p igf 1r rabbit antibody  (Cell Signaling Technology Inc)

Journal: Aging Cell

Article Title: Hormone replacement therapy enhances IGF-1 signaling in skeletal muscle by diminishing miR-182 and miR-223 expressions: a study on postmenopausal monozygotic twin pairs

doi: 10.1111/acel.12245

Figure Lengend Snippet: The effects of estrogen stimulation on the expression of miR-182, miR-223, and miR-142-3p and their targets in infant female quadriceps femoris -derived myoblasts. (A) Quantitative PCR analyses of miRNA transcripts normalized with RNU44 in human myoblasts treated for 72 h with 100 n m estradiol or mock. Note: The expression of miR-182 in these myoblasts was too low to be accurately measured. (B) qPCR analyses of target mRNAs (IGF-1, FOXO3A, FOXO1A) in human myoblasts treated with 10 n m and 100 n m estradiol or mock for 72 h. (C) Representative Western blots of target proteins, IGF-1R, FOXO3A and FOXO1A, and GAPDH, in human myoblasts treated with 10 n m and 100 n m estradiol or mock for 72 h. (D) Densitometry data of Western blots normalized with GAPDH. (E) Representative Western blots showing phosphorylation of AKT and mTOR proteins in myoblasts treated with 10 n m and 100 n m estradiol or mock for 72 h. (F) Densitometry data of Western blots normalized with GAPDH. Data are presented as percentage of control (mock) and reported as means ± SD of three independent experiments. OD indicates optical density. t -test, *** P < 0.001 , ** P < 0.01, * P < 0.05.

Article Snippet: For the MCF-7 cell and mouse muscle experiments, membranes were blocked in TBS with 0.1% Tween-20 (TBS-T) containing 5% fat-free dry milk for 60 min and then incubated overnight at 4 °C with rabbit polyclonal primary antibodies against FOXO1A, FOXO3A, IGF-1R, p-AKT, and total AKT diluted 1:1000 (Cell Signalling Technology) and against β-actin diluted 1:10000 (Santa Cruz, CA), which was used to check the uniformity of blotting.

Techniques: Expressing, Derivative Assay, Real-time Polymerase Chain Reaction, Western Blot, Phospho-proteomics, Control