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. Author manuscript; available in PMC: 2017 Sep 26.
Published in final edited form as: Cell Rep. 2017 Apr 4;19(1):101–113. doi: 10.1016/j.celrep.2017.03.038

Figure 4. Tumor Cell MAPK Signaling Is Critical for Viability following Taxol Treatment.

Figure 4

(A) Analysis of ERK1/2 signaling in tumor cells in response to treatment with Taxol (50 nM) in the presence or absence of BMDM-CM supplemented daily [denoted BMDM-CM(+)]. Representative western blots are shown. Area under the curve (AUC) quantification of band intensities is displayed in the graph below as a ratio of phosphorylated/total ERK1/2 (n = 3 independent experiments).

(B) Flow cytometric analysis of sub-2C DNA content tumor cells in response to Taxol (50 nM) and the MEK inhibitor PD0325901 (500 nM). Tumor cells were treated alone or in co-culture with BMDMs (n = 3 independent experiments).

(C) Analysis of ERK1/2 signaling and caspase-3 activation in tumor cells following 48 hr of treatment with Taxol (50 nM) in the presence or absence of BMDM-CM(+) and the MEK inhibitor PD0325901 (500 nM). Representative western blots are shown. AUC quantification of band intensities is displayed to the right as a ratio of CC3+/GAPDH (n = 3 independent experiments).

(D) Clonogenic assay measuring the effects of the MEK1/2 inhibitor PD0325901 and BMDM-CM on modulating inhibition of tumor cell colony formation by Taxol. Representative images are shown on the left, and quantification of crystal violet staining is shown on the right (n = 4 independent experiments). Scale bar, 5 mm.

(E) Tumor volume plots for the assessment of response to a single dose of Taxol (indicated by vertical dotted line) and acute BLZ945 or PD0325901 treatment. See Figure S5E for schematic of trial design. n = 10 mice in all treatment groups.

(F) Tumor volume plots for the assessment of response to weekly Taxol treatment (indicated by vertical dotted lines) with concurrent PD0325901 or sustained BLZ945 treatment. See Figure S6B for schematic of trial design. n = 10 mice in all treatment groups.

All data are presented as mean ± SEM. Student’s t test was used to assess significance between treatment groups: *p < 0.05; **p < 0.01; ***p < 0.001. See also Figures S5 and S6.