Fig. 5

To stimulate tumor cell repopulation, Hsp70-HMGB1 complex is needed, not Hsp70 or HMGB1 separately. (A) Western blot of HBHP- and HMGB1-scr peptide eluates from Eto-CM of A549 cells. (B) A549 cells were seeded into wells of 96-well plates at a low concentration and then were incubated in Eto-CM, full and depleted with HBHP-peptide as well as HBHP-scr peptide. Cell growth was evaluated with MTT assay on day 14. **p < 0.001.(C) Level of PGE2 expression in in full or depleted Eto-CM measured with use of human PGE2 immunoenzyme assay kit; (D) HMGB1 was depleted from CM of untreated A549 cells and Eto-CM and then purified HMGB1 or purified Hsp70, or their complex were added and these media and were applied to A549 seeded to E-plates at a low concentration. Cell growth in real time was estimated using the xCELLigence technique. (E) A549 cells were incubated with the same CMs as in (C) and (D) for 24 h and then cells were lysed and used for western blotting with antibodies against autophagy marker LC3 I-II