Fig. 5.

a TEM images of HaCaT cells. Left panel: untreated, size bar 1 μm; right: α-toxin for 2 h, size bar 500 nm. b Digital fluorescence microscopic image of HaCaT cells treated with AlexaFluor546-labelled α-toxin (red) for 2 h. Left image sample was fixed and stained for p62 (green), by using AlexaFluor488-coupled secondary antibody. The image of the right panel was taken from cells which were transiently transfected with EGFP-LC3, and treated with AlexaFluor546-α-tox as above. Arrows indicate colocalization of AlexaFluor546-α-tox with p62, or EGFP-LC3-positive puncta, respectively. Untreated controls exhibited diffuse distribution of p62 (not shown), and of most EGFP-LC3 (see Fig. 6a)