Phospho p70 s6k t421 s424

NIH 3T3 fibroblasts were obtained from ATCC (Manassas, VA) and maintained in DMEM (HyClone, Logan, UT) supplemented with 10% fetal bovine serum, 100 units/ml penicillin, and 100 µg/ml streptomycin in a 5% CO₂ incubator at 37°C. Primary antibodies against phospho-Akt^S473, phospho-Mek^S217/221, Pan-Mek, phospho-S6 ribosomal protein^235/236, phosphop-70S6K^T421/S424, phospho-ERK1/2^42/44, phospho-4EBP^37/46 and pan-Akt were purchased from Cell Signaling (Boston, MA). Primary antibodies against phospho-p70S6K^Ser411 were purchased from Abcam (Cambridge, MA). Bleomycin and primary antibodies against β-actin and fibronectin were purchased from Sigma (St. Louis, MO). Anti-collagen-I was purchased from Rockland (Gilbertsville, PA). All HRP-conjugated secondary antibodies were obtained from Bio-Rad (Hercules, CA) and Alexa-labelled secondary antibodies were purchased from Life Technologies (Carlsbad, CA). TGFβ was purchased from R&D (Minneapolis, MN), whereas LY294002 inhibitor, U0126 inhibitor, and Rapamycin were purchased from EMD Millipore Bioscience (San Diego, CA). PF4708671 inhibitor was obtained from Santa Cruz Biotechnology (Dallas, TX). Adenoviral particles encoding CA-Mek1 and DN-Mek1 were obtained from Cell Biolabs, Inc. (San Diego, CA), whereas retroviral plasmids encoding CA-Akt1 and DN-Akt1 were generated in the laboratory [6 (link)].