Figure 4. Combination treatment of GNF5 and docetaxel induces lung tumor cell differentiation in vivo.

GFP+ cells were isolated from KRAS^LSL-G12D; p53^fl/fl; Rosa26-fGFP mice two weeks after treatment with vehicle, docetaxel, GNF5, or combination treatment and 12 weeks after induction of tumors with adenovirus. A. Quantification of total tumor volume in bilateral lungs evaluated by μ-CT scans taken before (10 weeks) and after (12 weeks) treatment with vehicle or combination therapy (GNF5 + docetaxel) shows that combination treatment significantly impairs tumor growth in mice compared to vehicle treated mice (n = 11 mice per group). B. Western blot analysis showed an increase in expression of the ciliated cell marker, acetylated α-tubulin, and the secretory cell marker, Scgb1a1, with a corresponding decrease in expression of the basal cell marker, keratin 5, in mice treated with GNF5 and docetaxel. C. RT-qPCR analysis for indicated cell treatment groups shows an increase in expression of terminal cell markers (Sftpc: Type II cell marker and Scgb1a1: secretory cell marker) with a corresponding decrease in expression of the basal cell marker, TP63, in mice treated with the combination therapy compared to control or mice treated with GNF5 or docetaxel alone (n = 4-6 mice per group, each RT-PCR assay performed in triplicate). Graphs depict means and S.E.M.