Fig. 2

(A) Densitometric evaluation of relative pixel intensities (normalized to untreated controls) of the indicated phosphorylation sites in cell lysates from GCT72, 1411H, TCam-2 and BeWo cells treated with recombinant CXCL12 (250 ng / ml) for 24 h, as measured by a human phospho-kinase array. (B) Relative migration of GCT72 and 1411H cells treated with either 100 ng / ml recombinant CXCL12, 100 nM CXCR7-NB (VUN702), or the combination of both, in comparison with the untreated control. (C) Box plot summarizing the number of proliferative GCT72 cells treated with 100 ng / ml recombinant CXCL12, 20 µM CXCR4-inhibitor AMD3100, or the combination of both for 24 h in comparison to the untreated control. (D) Box plot summarizing the number of proliferative 1411H cells treated with 100 ng / ml recombinant CXCL12, CXCR4-inhibitors WZ811 (5 µM) / LY2510924 (50 nM), or the combination of both for 32 h in comparison to the untreated control. (E) Densitometric evaluation of western blot data of phospho- and total-ERK in EC cells (2102EP, NCCIT, NT2/D1) treated daily with 100 nM ERK inhibitor SCH772984 for 96 h. (F) Relative mRNA expression of CXCR4 and CXCR7 in EC cell lines (2102EP, NCCIT, NT2/D1) treated daily with 100 nM SCH772984 for 96 h as compared to untreated controls. ACTB and GAPDH were used as housekeeping genes. (G) Model summarizing key findings related to the CXCR4 / CXCR7 / CXCL12 axis. SEM present as CXCR4⁺ / CXCR7⁻, YST as CXCR4⁺ / CXCR7⁺, CC as CXCR4⁻ / CXCR7⁺ and EC as CXCR4⁻ / CXCR7⁻. SEM (also with occult YST subpopulations), YST and CC are targetable by CXCR4- and / or CXCR7-NDC, respectively. CXCL12 stimulated CXCR4 / CXCR7 enhanced proliferation and migration in YST cells. In EC cell lines, inhibition of MAPK (ERK1 / 2) signaling allows for re-induction of CXCR7 expression (and partly CXCR4)