Extended Data Fig. 6: Glutaminolysis is essential for post-transcriptional regulation of actin dynamics and efferocytosis.
(a) Schematic representation of efferocytosis steps and RNAseq analysis of “find-me”, “eat-me” and “tolerate-me” signals in reparative IL-4 treated control and MacΔGls1 PCMs. (b) Schematic representation of actin polymerization and depolarization (left panel) and RNAseq analysis with focus on F-actin dynamic regulators in reparative control or Mac∆Gls1 PCMs (right panel). (c) Dbl expression quantified by flow cytometry in control or IL-4-treated macrophages that were incubated ± ACs for 45 min. (d) RNAseq analysis with focus on GTP-converting ezymes. (e) qPCR quantification (left) and efferocytic index (right) in Gls1fl/fl control and Mac∆Gls1 BMDMs stimulated overnight with IL-4 after Nme1/6 or Sucgl1 lentivirus overexpression. (f) Cdc42 and (g) Rac1 activity assays in control or Mac∆Gls1 BMDMs in basal condition or stimulated overnight with IL-4 + /− AOA. (h) Efferocytic index in reparative control or Mac∆Gls1 BMDMs or (i) in resolving control or Mac∆Gls1 BMDMs + /− NSC23766 (RAC1 inhibitor), ML141 (CDC42 inhibitor) or MDIVI-1 (DRP1-mediated mitochondrial fission inhibitor). All values are mean ± SEM and are representative of at least one independent experiment (n = 2 to 3 independent animals for c-f, n = 5 to 9 for g, n = 3 to 6 for h, i). P values were determined by ordinary one-way ANOVA with Tukey post hoc test for multiple comparisons (c, e-i). Each statistical bar color-coded represents an independent one-way ANOVA test. Source data are provided as a Source Data file (c, e-i).
