Figure 5
From: Partial loss of Smad signaling during in vitroprogression of HPV16-immortalized human keratinocytes
Western blot analysis of Smad2 phosphorylation in normal HKc, HKc/HPV16, and HKc/DR after 6 h of TGF-β treatment. Normal HKc, low-passage (< 40) HKc/HPV16, and HKc/DR were treated for 6 h with 40 pM TGF-β. Whole-cell extracts were collected and total protein (25 μg/well) was resolved in a 10% polyacrylamide gel. A rabbit anti-phospho-Smad2 antibody, a mouse anti-Smad2 monoclonal antibody, and a rabbit anti-actin polyclonal antibody were utilized to detect phospho-Smad2, total Smad2, and actin, respectively. Panel A shows two normal HKc strains, and HKc/HPV16 and HKc/DR from two donors (d1 and d2). Panel B shows the results from two additional normal HKc strains, as well as HKc/HPV16 and HKc/DR from two additional donors (d4 and d5).