Fig. 4.

Autophagy influences the sensitivity of B16 cells to CTLs. a, b Autophagy affects the cytolysis to B16 cells. B16 cells (2 × 10⁵) were inoculated to mice. 12 days later, spleen T cells were isolated and stimulated in vitro with HSP70-B16 peptide complexes in the presence of irradiated normal splenocytes as APCs for 6 days. The cells were harvested as effector cells. B16 cells were treated with rapamycin, 3-MA, BM, wortmannin, or chloroquine in the presence (a) or absence (b) of IFN-γ for 24 h, and used as target cells. The cytolysis assay was performed as described in “Materials and methods”. *P < 0.05, compared with PBS group (20:1). ^# P < 0.05, compared with PBS group (10:1). c siRNA silences Beclin 1 expression. After stable transfection of Beclin 1 siRNA vector into the B16 cell line, Beclin 1 mRNA and protein expression were examined by RT-PCR and Western blot. d Beclin 1 knockdown affects the cytolysis to B16 cells. The above effector cells were generated. Beclin 1 siRNA-expressing B16 cells were treated with or without IFN-γ for 24 h, and used as target cells. The cytolysis assay was performed (*P < 0.05)