The Putative HORMA Domain Protein Atg101 Dimerizes and Is Required for Starvation-Induced and Selective Autophagy in Drosophila
Figure 2
RNAi depletion of Atg101 by expression of a long hairpin (a) or a microRNA-based silencing construct (b) in clones of cells marked by membrane-bound mCD8-GFP prevents starvation-induced punctate LysoTracker Red (LTR) staining, compared to surrounding control fat body cells of third instar larvae. (c) Quantification of data shown in (a) and (b), , , two-tailed, two-sample Student’s t-tests. (d) Reverse Transcriptase-PCR analysis reveals that Atg101 mRNA expression is strongly reduced upon expression of Atg101[KK], and it is undetectable in Atg101[HMS] expressing animals compared to wild type L3 stage larvae. Note that the expression level of Atg101 is much lower than that of Actin5C, a commonly used control in such experiments. Numbers in parentheses indicate the amplified region of the coding sequence of these genes. Scalebar equals 20 μm for microscopic images.
