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. 2014 Sep 16;5(23):12027–12042. doi: 10.18632/oncotarget.2482

Figure 7. Macrophages stimulate melanosphere formation via TGFβ.

Figure 7

For panels (B) to (E), data are expressed as fold change in sphere-forming efficiency relative to untreated CD34− TICs. Statistical analysis: * P<0.05, ** P<0.01, *** P<0.001, **** P < 0.0001. (A) Graph comparing the Log2 expression of the tgfb gene relative to GAPDH in TAMs and CD34− TICs. Bars represent mean ± SD with each point representing cells from one mouse, two-tailed Mann-Whitney test. (B) Effect of TGFβ inhibitors on the stimulatory effect of TAMs: Anti-TGFβ antibody (1μg/ml) and SD208 (1μM) were added to CD34− TIC cultures with or without TAMs for the duration of the experiment. Bars represent mean ± SE, 2-way ANOVA. (C) Effect of TGFβ inhibitors on the stimulatory effect of M2-CM: Anti-TGFβ and SD208 were either added to the BMDM prior to CM generation (SD208 > CM, α-TGFβ > CM) or added to the CM during sphere culture (CM+SD208, CM+α-TGFβ). Bars represent mean ± SE. Line above bars represents comparison between M2-CM and each of the treatments, 1-way ANOVA. (D) Graph showing the dose-dependent stimulatory effect of TGFβ on sphere formation. Bars represent mean ± SE. Line above bars represent analysis of linear trend, 1-way ANOVA. (E) Effect of M2-BMDM derived from tumor-bearing mice (RET+), wild-type mice (WT) and Smad3 knockout mice (SMAD3KO) on sphere formation. Bars represent mean ± SE, 1-way ANOVA. (F) Comparison of Arg1 activity within WT and SMAD3KO M2-BMDM reflected by the amount of urea production. Bars represent mean ± SD, two-tailed Mann-Whitney test.