Fig. 1

Autophagy activation by PFD in LF. a Fluorescence microscopic detection of pEGFP-LC3 dot formation in LF: LF were transfected with pEGFP-LC3 and control siRNA, ATG5 siRNA, or ATG7 siRNA. Photomicrographs are taken at the same magnification. (Original magnification, 200X). The lower panel is the percentage of positive cells with more than five dot formations (±SEM) and data was collected from four independent experiments. *p < 0.05. b Western blotting (WB) using anti-LC3 and anti-β-actin of cell lysates from control (lane 1, 2), PFD (500 μg/ml) (lane 3, 4) treated LF. LF were treated with PFD for 24 h and protease inhibitor (E64d 10 μg/ml, pepstatin A 10 μg/ml) treatment was started 6 h before collecting cell lysates. In the lower panel is the average (±SEM) taken from six independent experiments shown as relative expression. *p < 0.05. c WB using anti-LC3 and anti-β-actin of cell lysates from control siRNA (lane 1 ~ 4), ATG5 siRNA (lane 5 ~ 8) transfected LF. PFD (500 μg/ml) treatment was started 48 h post transfection and protein samples were collected after 24 h treatment. Protease inhibitor (E64d 10 μg/ml, pepstatin A 10 μg/ml) treatment was started 6 h before collecting cell lysates. In the right panel is the average (±SEM) taken from five independent experiments shown as relative expression. *p < 0.05. d WB using anti-LC3 and anti-β-actin of cell lysates from control siRNA (lane 1 ~ 4), ATG7 siRNA (lane 5 ~ 8) transfected LF. PFD(500 μg/ml) treatment was started 48 h post transfection and protein samples were collected after 24 h treatment. Protease inhibitor (E64d 10 μg/ml, pepstatin A 10 μg/ml) treatment was started 6 h before collecting cell lysates. In the right panel is the average (±SEM) taken from five independent experiments shown as relative expression. *p < 0.05