Figure 7. The volasertib acquired resistance (VAR) cell line shows mesenchymal‐to‐epithelial transition and persistent cMet phosphorylation.

• A
  Cell viability measured by CellTiter‐Glo in Calu6 parental and Calu6‐VAR cells treated with different concentrations of volasertib for 72 h.
• B
  Micrograph shows acquisition of a mesenchymal‐to‐epithelial transition phenotype in VAR cells. Scale bar: 100 μm.
• C
  Basal protein and mRNA expression in parental and VAR cells for the indicated proteins was measured using immunoblotting (upper) and reverse‐transcription PCR (lower), with GAPDH expression used for normalization.
• D
  Parental and VAR cells were treated as indicated for 48 h, and apoptosis was measured using the Apo‐BrdU assay.
• E
  Parental and VAR cells were treated with the indicated drug concentrations for 48 h. Cells were then harvested, and lysates were immunoblotted for the indicated proteins.
• F
  Parental and VAR cells were treated as indicated for 24 h and allowed to grow in drug‐free medium for 15–20 days to form colonies, which were counted using ImageJ.

Data information: Data are means ± standard error of the mean from three independent experiments. Significant differences using two‐way analysis of variance with Bonferroni or Benjamini–Hochberg correction for multiple comparisons are indicated. *P < 0.01.Source data are available online for this figure.