Fig. 5: CP competitively interacted with PEBP4 to liberate ATG8 and promoted autophagy. | Nature Communications

Fig. 5: CP competitively interacted with PEBP4 to liberate ATG8 and promoted autophagy.

From: PEBP balances apoptosis and autophagy in whitefly upon arbovirus infection

Fig. 5

a GST pull-down showing the interaction between recombinant His-ATG8 and GST-PEBP4. b PEBP4 co-immunoprecipitated with ATG8 in viruliferous (V) or nonviruliferous (N) whitefly. c–e Recombinant His-CP and GST-PEBP4 with concentration gradients were co-incubated with the lysate of whitefly to monitor the activation of autophagy by immunoblotting, PEBP4 bands represent endogenous abundance. f Competition between CP and ATG8 for PEPB4 by competitive pull-down assays. His-CP and His-ATG8 were supplied within a concentration gradient to co-incubate with GST-PEBP4. g ATG8 (blue), PEBP4 (red), and CP (green) were simultaneously labeled and localized by immunofluorescence in viruliferous whitefly. The nuclei were stained by DAPI (white). Sample sizes (n) for statistical tests indicated in the panels refer to biologically independent whitefly. Colocalization was quantified by ImageJ Coloc2 using Manders’ Colocalization Coefficients (MCC). M1 = CP-ATG8/Total ATG8; M2 = CP-ATG8/Total CP; M3 = PEBP4-ATG8/Total ATG8; M4 = CP-PEBP4/Toal CP. h Model of CP-PEBP4 regulated autophagy in whitefly. Unconjugated ATG8 arrested PEBP4 to prevent autophagy in nonviruliferous whitefly. Upon TYLCV invasion, CP competed for PEBP4 binding, which released ATG8 and initiated autophagy.

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