(A) Schematic diagram of WT and mutant TAX1BP1 constructs.
(B) Representative CFSE dilution histograms of stimulated Tax1bp1−/− CD4+ T cells transduced with the indicated TAX1BP1 mutants, rested, and subsequently re-stimulated with anti-CD3 and anti-CD28 antibodies for 2 days (upper panel). Quantitation of cells ≥2 cell divisions shown (lower panel). Mean values ± SD. **p < 0.01 by one-way ANOVA with Sidak’s test.
(C and D) Immunoblot analysis of TAX1BP1 constructs expressed in 293 T cells (left panel) (C and D) and their interaction with LC3B (right panel) (C) and their interaction with ubiquitin (right panel) (D).
(E) Endogenous TAX1BP1 and p62 interactions with LC3 family proteins. Whole cell lysate from activated wild type CD4+ T cells was harvested, precipitated with the indicated GST fusion proteins of LC3 family members, and immunoblotted for TAX1BP1 or p62 expression.