Fig. 3

The repopulation-stimulating effects of CM is not exhibited by Hsp70 alone, but by Hsp70 together with some binding proteins. (A) A scheme demonstrating the difference in the composition of eluates during ATP precipitation and immunoprecipitation. (B) Ag-stained gel from CM from untreated A549 cells and from Eto-CM and Oxa-CM performed with specific antibodies and with ATP-agarose. (C) A549 and DLD1 cells were seeded into wells of 96-well plate at a concentration of 500 cells per well and CMs from untreated eponymous cells of Eto-CM and Oxa-CM depleted with ATP or IP were applied for 14 days, then cell growth was evaluated using MTT test. ** p < 0.001. (D) The A549 cells were treated as in (C) and their growth was evaluated with xCELLigence technique in real time. The results of one of many experiments are presented. Data for DLD1 cells, see the supplementary file Suppl (Fig. 2S). (E) Colony formation of A549 and DLD1 cells in presence of Eto-CM or Oxa-CM depleted with ATP or IP. (F) Colonies intensity, presented in folds of control. (G) Expression of repopulation marker genes in A549 and DLD1 cells in presence of Eto-CM or Oxa-CM depleted with ATP or IP