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. 2019 Apr 10;5(4):eaau8237. doi: 10.1126/sciadv.aau8237

Fig. 2. Mislocalization and altered conformational status of GABARAPs in Atg7 cKO brains.

Fig. 2

(A) Schematic diagram of SDD-AGE/SILAM experiment. (B) Abundance of proteins identified in high molecular weight fraction (>250 kDa) only in CaMKII-cre Atg7 cKO but not control brains by SDD-AGE/SILAM experiments (6-month-old mice). (C) Candidate proteins meeting the selection criteria are listed with SILAM scores representing relative abundance to SILAM-labeled sample. Selection criteria are depicted schematically in fig. S2B. Candidate proteins identified by these criteria at all three ages examined are specifically marked by asterisks. (D) GABARAPL2 and p62 were consistently identified by SDD-AGE/SILAM in CaMKII-cre Atg7 cKO brain homogenates at 3 months (left) and 12 months (right) of age. (E) p62-like accumulation of GABARAPs was observed in Dlx5-creERT2 and CaMKII-cre Atg7 cKO mice. (F) Native PAGE analysis revealed shifts of GABARAPL2 and GABARAP into high–molecular weight (MW) complexes in the brains of CaMKII-cre Atg7 cKO mice (cortex). (G to I) GABARAPs were observed to mislocalize to p62+ aggregates in affected neurons of Dlx5-creERT2 Atg7 cKO mice. Scale bars, 20 μm (G to I).