Fig. 4: Analysis of HGF/met signaling and stemness/adhesion-associated markers.

a Western blot analyses subsequent to growth of the indicated cell populations in the absence of FCS for 24 h. The c-met antibody detects the unprocessed single chain precursor protein (upper band, 170 kDa) as well as the mature 140 kDa β-chain (lower band). The phospho-met antibody detects phosphorylated Tyr1234/Tyr1235 residues in the c-met β-chain. ACTB was probed as a control for equal protein loading, respectively. b Cell surface integrin profiling at 72 h post target gene induction using flow cytometry of live cells. Representative histogram plots for one of two experimental series showing similar results are depicted. Cell populations are color coded as indicated in the legend. “VC unstained” denotes PC3L1-VC cells which were incubated with the fluorophore coupled secondary antibody only, omitting the antigen-specific primary antibody. c Exemplary western blot analysis of CD49f/ITGA6 resulted in the detection of glycosylation variants at ~125 and 150 kDa. ACTB was probed as a control for equal protein loading. d Representative two parameter dot plots showing expression levels of CD24 (horizontal axis) vs. CD44 (vertical axis) in double-staining experiments. Cell populations were gated for live cells and subjected to quadrant analysis using identical settings. One of two experimental series providing similar results is shown.