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. Author manuscript; available in PMC: 2018 Sep 15.
Published in final edited form as: Cancer Res. 2017 Jul 7;77(18):5142–5157. doi: 10.1158/0008-5472.CAN-16-2586

Figure 3. CAF-MSC and BM-MSC promote tumor cell viability by increasing proliferation and inhibiting apoptosis.

Figure 3

A: Indicated NB-FLuc cells were cultured in direct contact with CAF-MSC, BM-MSC or Fb (ratio 4:1), and examined for viability over 4 days by adding D-Luciferin as described in Materials and Methods. As controls, NB cells were grown alone in their own conditioned medium. The data represent the mean (±SD) luminescence activity in relative light units (RLU) from six samples and are representative of one of three independent experiments showing similar results; B: Indicated NB cells were co-cultured in a transwell system using the same conditions as in A. The data represent the mean number (±SD) of trypan blue-negative cells in triplicate samples and are representative of one of two independent experiments showing similar results; C: Indicated NB cells were cultured in CM from NB, NB/CAF-MSC, NB/BM-MSC or NB/Fb co-cultures and examined for viability by CellTiterGlo as described in Materials and Methods, over 4 days as in A. The data represent the mean (±SD) luminescence activity of six samples and are representative of one of two independent experiments; D: Indicated NB cells were cultured as described in C and examined for BrdU incorporation after 48 hours as indicated in Materials and Methods. The histograms represent the mean (±SD) percentage of cells in S-phase for triplicate samples and results are representative of one of three independent experiments showing similar results; E: NB cells were cultured as indicated in C and examined for apoptosis after 4 days by flow cytometry as indicated in Materials and Methods. The histograms represent the mean (±SD) percentage of Annexin V+/PI+ and Annexin V+/PI− cells for triplicate samples, and results are representative of one of two independent experiments showing similar results. *p<0.05;**p< 0.01; ***p<0.001; ****p< 0.0001; n.s. = not significant.