Figure 3

rCST9 induced autophagy. (A) MDM showed evidence of autophagy (that is, double membrane AV) in MDM infected with S4 (MOI 40:1) at 20 h after rCST9 (50 pg) treatment and S4 infection as per TEM analysis. (B) Immunofluorescent microscopy showed LC3-II protein (small green spots) in human alveolar macrophages at 20 h after rCST9 (50 pg) treatment compared with untreated macrophages. Images shown in A and B are representative of experiments repeated three times using duplicate conditions. (C) Western blot analysis revealed marked conversion of LC3-I to LC3-II in rCST9 (50 pg) treated MDM compared with MDM alone. (D) rCST9 (50 pg) induced the dephosphorylation of P70S6 kinase at 30 and 60 min after treatment. PI3-KC3 phosphorylation of was detected at 10, 30 and 60 min as well as 4, 5 and 6 h in rCST9-treated MDM. Experiments were performed three times.