Fig. 6

Inhibition of the thioredoxin-reducing system or oxLDL treatment prevents LC3 lipidation. a The overexpression of TXNIP in HEK cells but not EGFP significantly attenuates LC3 lipidation in response to amino acid withdrawal, observed in the presence or absence of bafilomycin A1. The graph below shows quantification of LC3 lipidation from treated EGFP or TXNIP transfected HEK cells. b During amino acid withdrawal the loss in LC3 interaction with Atg3 and Atg7 is maintained in cells overexpressing TXNIP or EGFP. The graphs below show quantification of Atg3-LC3 and Atg7-LC3 formation from treated EGFP or TXNIP transfected HEK cells. c Treatment of SMC with oxLDL significantly attenuates basal LC3 lipidation and leads to accumulation of LC3I. The graphs to the right show quantification of LC3I and LC3II from control, EBSS or oxLDL-treated SMC. d oxLDL induces Atg7 oxidation and significantly decreases the covalent interaction between Atg3 and LC3. The graphs to the right show quantification of Atg7 oxidation, Atg3-LC3 and Atg7-LC3 formation from control, EBSS or oxLDL-treated SMC. All data represent mean ± s.e. from 4−5 (a, b) or 3 independent experiments (c, d). *P < 0.05, **P < 0.01, ***P < 0.005 statistical significance (Dunnett’s test) relative to control or bafilomycin A1 alone. ††P < 0.01, †††P < 0.005 statistical significance relative to EBSS or EBSS + bafilomycin A1 treatment