Fig. 9: Rubicon lactylation promotes LAP activation.
From: PDCD6 regulates lactate metabolism to modulate LC3-associated phagocytosis and antibacterial defense
a, b Immunoblotting of total lactylation in Pdcd6fl/fl and Pdcd6Δmye BMDMs treated or not with L. monocytogenes (a) or Pam3CSK4 (b) for the indicated periods. c–e Total Kla (c) and Rubicon (d, e) were immunoprecipitated from Pdcd6fl/fl, and Pdcd6Δmye BMDMs challenged or not with L. monocytogenes (MOI: 5) (c, d) or zymosan (e), followed by immunoblotting with the indicated antibodies. f, g Lactylation mapping in RUBCN truncation mutants (f) or RUBCN K-to-M mutants (aa 1–180) (g) by immunoprecipitation using HEK293T cells. h Reconstitution of THP-1 cells with RUBCN WT or RUBCN K33R mutant. i–l Gentamicin protection assays (i), flow cytometric analysis of LC3B fluorescence (j), and colocalization analysis between LC3B and RFP-L. monocytogenes (k, l) in THP-1 cells with the reconstitution of either RUBCN WT or the RUBCN K33R mutant. Scale bar, 2 µm. m Cross-species sequence alignment of RUBCN highlighted the conservation of the K33 lactylation site. The averages of n = 4 (i), n = 3 (j), and n = 10 (l) biologically independent samples are shown. Data are shown as the mean ± SEM. Statistical significance in (i, j, l) was determined using the t test (and nonparametric tests). The data presented in (a–k) are representative of three independent experiments.
