Fig. 10.
Analysis of Atg4D localisation in response to cellular stress. HeLa cells transiently expressing GFP-Atg4D were starved (A), treated with staurosporine (1 μM) (B) or treated with hydrogen peroxide (10 nM) (C) for 5 hours, then fixed and labelled with antibodies against HSP60. In a sub-population of starved cells, small GFP-Atg4D puncta were observed in the vicinity of mitochondria (Aii); however, no evidence of mitochondrial Atg4D localisation was observed in starved or staurosporine-treated cells (A,B). Note that in late apoptotic cells Atg4D formed aggregates, but these did not colocalise with mitochondria (Bii; see also supplementary material Movie 1). (C) GFP-Atg4D relocates to mitochondria in a sub-population of H2O2-treated HeLa cells. Mitochondria not labelled with GFP-Atg4D are indicated with arrows. In parts (A) and (B) zoomed panels are shown to the right. Scale bars: 10 μm (A,B), 5 μm (C).