Extended Data Fig. 1: M45 induced aggregates.
(a) Atg5-/- MEFs were transfected with an M45-mCherry plasmid. 24 h post transfection cells were fixed and analyzed by fluorescence microscopy. Nuclei were stained with Hoechst 33342. Scale bar, 10 µm. (b) NIH-3T3 cells transfected with plasmids expressing HA-tagged full-length M45 or M45-Ct3. HA-tagged proteins were detected by immunofluorescence (green), protein aggregates by using the ProteoStat dye (red). (c) Another two ultrathin sections of the same WT MEF cell shown in Fig. 1f (ix – xi). The arrow points to a different aggregate than the one in Fig. 1f. Magnification of the same aggregate (A) close to an autophagosome (*) in the two different sections (x – xii). Scale bars, 5 µm (ix, xi) and 200 nm (x, xii). (d) Maximum intensity projection of NIH-3T3 cell transfected with a plasmid encoding M45-mCherry and observed 6 h post transfection by live cell imaging for 30 h. Scale bar, 10 µm. These data are representative of three (a-b) or two (d) biologically independent experiments. This data (c) is representative of three independent infected cells per group where at least 10 different sections (50 nm) per cells were analyzed.
