Fig. 3: HMGB1 involved in DHA-37-induced autophagic cell death in A549 cells.

a A549 cells were treated with DHA-37 at indicated concentrations for 24 h before cell lysis. Cell extracts were analyzed by western blotting. b A549 cells were treated with 10 μM HA-37 at indicated time before cell lysis. Cell extracts were analyzed by western blotting. c A549 cells were treated with DHA-37 at indicated concentrations for 24 h and total RNA was extracted, mRNA levels were detected using quantitative polymerase chain reaction (Q-PCR). d, e A549 cells were transfected with control siRNA or siRNA target HMGB1. The knockdown of HMGB1 was confrmed by western blot (d). The cells were treated or not with 10 μM DHA-37 for 24 h. Cell extracts were analyzed by western blotting (e). f A549 cells were transfected or not with siRNA targeting HMGB1 and control siRNA, respectively. After 24 h, cells were treated with 10 μM DHA-37 for 48 h, and cell viability was measured by MTT assay. g, h A549 cells were transfected with pcDNA3.1-HMGB1 or pcDNA3.1 for 24 h, respectively, Cells were treated or not with 10 μM DHA-37 for 24 h, the cell extracts were analyzed by western blotting. i A549 cells were transfected or not with HMGB1 over-expression vector and empty vector, respectively. After 24 h, cells were treated with 10 μM DHA-37 for 48 h, and cell viability was measured by MTT assay. The data are presented as the mean ± SD or SEM. n ≧ 3. *p < 0.05, **p < 0.01