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. Author manuscript; available in PMC: 2020 Jul 13.
Published in final edited form as: Nat Cell Biol. 2020 Jan 13;22(2):187–199. doi: 10.1038/s41556-019-0450-y

Extended Data Fig. 5: LC3-conjugation machinery controls EV-mediated secretion of diverse RBPs.

Extended Data Fig. 5:

a, EVs from WT and ATG deficient cells normalized for protein concentration and immunoblotted to detect endogenous LC3A, LC3B, LC3C, GABARAP (GR), GABARAPL1 (GRL1), GABARAPL2 (GRL2), and indicated marker proteins (n=2 biologically independent experiments). b, Whole cell (WCL) and EV lysates from WT and ATG7−/− cells were normalized for protein concentration and immunoblotted for indicated proteins (n=3 biologically independent experiments). c, HEK293T cells co-transfected with FLAG-tagged G3BP1, LARP1 or SF3A1, and myc-tagged LC3B, GABARAP (GR), LC3C respectively, or myc-BirA* were lysed, immunoprecipitated (IP) with anti-myc antibody and immunoblotted (WB) with indicated antibodies (n=3 biologically independent experiments). d, Diagram mapping the domains and primary LC3-interaction region (LIR) in SAFB. e, Volcano plot of mRNA and long non-coding RNA (large RNA) detected in EVs from WT and ATG7−/− cells. Results plotted according to -log10 p-values as determined by DESeq2 and log2 fold enrichment (n=3 biologically independent samples; WT/ATG7−/−). Grey dots: RNAs not enriched in EVs from WT or ATG7−/− cells identified with a p-value >0.05 and/or log2 fold change between −0.5 and 0.5 (−0.5<log2FC<0.5). Black dots: Large RNAs enriched in EVs from WT cells or ATG7−/− cells. f, Volcano plot of mRNA and long non-coding RNA (large RNA) detected in EVs from WT and ATG12−/− cells. Results plotted according to -log10 p-values as determined by DESeq2 and log2 fold enrichment (n=3 biologically independent samples; WT/ATG12−/−). Grey dots: RNAs not enriched in EVs from WT or ATG12−/− cells identified with a p-value >0.05 and/or log2 fold change between −0.5 and 0.5 (−0.5<log2FC<0.5). Black dots: Large RNAs enriched in EVs from WT or ATG12−/− cells. g, Venn diagram showing the overlap of mRNA and long non-coding RNAs (large RNAs) enriched in EVs from WT relative to ATG7−/− cells and EVs from WT relative to ATG12−/− cells. Data and unprocessed blots available in Source Data Extended Data Fig. 5.