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. 2016 Jul 28;7(35):56408–56421. doi: 10.18632/oncotarget.10895

Figure 2. The expression of CSF-1R and its ligand influence the resistance to cisplatin treatment.

Figure 2

(A) Cisplatin treatment increases mRNA and protein levels of CSF-1 and CSF-1R. Histograms reporting the mRNA levels of CSF-1 and CSF-1R, in cells harvested at 24 hr (quantitative PCR), and the protein levels of CSF-1 in medium conditioned for 48 hr (ELISA assay), from the indicated cell lines treated with cisplatin at the CC50 doses. (B) The CSF-1R expressing cells survive chemotherapy-induced stress. Upper panel. Representative FACS dot plots showing the percentage of CSF-1Rpos in H1299 cells treated with cisplatin at the CC50 and CC75 for 72 hrs (gated). Gated cells and isotype controls are further illustrated in Supplementary Figure S1B. Lower panel: histogram bars reporting the percentage of CSF-1Rpos cells in the indicated cell lines. In the lower panel, the mean ± SE of two independent experiments is reported. (C–D) CSF-1R inhibition affects the clonogenicity and resistance to cisplatin of lung cancer cell lines. (C) Western blotting of CSF-1R immunoprecipitates stained with the indicated antibodies from H1299 cells treated with increasing concentrations of JNJ-40346527. (D) Upper panel. Representative micrographs of the colonies formed by H1299 cells treated with increasing doses of JNJ-40346527. Lower panel. Histograms reporting the mean ± SE of the colonies formed by 4 representative cell lines in three independent experiments. (E) CSF-1R inhibition affects the resistance of lung cancer cell lines to cisplatin. Graphs reporting the percentage of colonies formed by A549 and H1299 cells treated with JNJ-40346527 at the CC25 doses determined in 2D and with the indicated doses of cisplatin. Mean ± SE of three independent experiments. (F) JNJ-40346527 treatment modulates the number of CSF-1Rpos cells. Histograms reporting the number of CSF-1Rpos cells (as assessed by FACS) in the H1299 cells treated for 96 hrs at the previously determined CC50 dosages for both cisplatin and JNJ-40346527. Mean ± SE of three independent experiments is reported. *= p < 0.05.