Extended Data Fig. 1: PERK deficiency does not affect M1 activation.
From: PERK is a critical metabolic hub for immunosuppressive function in macrophages
a, Enrichment plot of endoplasmic reticulum (ER) stress genes in IL-4c treated mouse peritoneal macrophages compared with naïve (PBS) macrophages by GSEA analysis. b, GSEA result comparing ER stress genes between TAMs and nontumor macrophages from patients with lung carcinoma. c, Pearson correlation of TAM CD68 expression with genes encoding molecules involved in the PERK pathway in caxncer patients from the TCGA. d, Immunoblot analysis of p-PERK, PERK, and β-actin from BMDMs stimulated with IL-4, Thapsigargin, or phosphatase. Ratio of p-PERK to total PERK was determined using ImageJ. Data are representative of three independent experiments. e, Frequency of p-PERK+ in BMDMs stimulated with IL-4 or Thapsigargin (n = 3; mean ± s.e.m). Data are collected from three independent experiments. f, Percentage of CD206+CD301+ in BMDMs treated with either DMSO (vehicle), IL-4, or Thapsigargin (n = 3; mean ± s.e.m). Data are collected from three independent experiments. g, Expression of CD206 and CD301 from BMDMs treated with IL-4 (M2) in the presence or absence of GSK2656157 (n = 3; mean ± s.e.m). Data are representative of three independent experiments. h, Percentage of 7AAD− naïve BMDMs isolated from Eif2ak3fl/fl or Eif2ak3fl/fl x LysMCre mice (n = 6; mean ± s.e.m). Data are collected from six independent experiments. i, Western blotting analysis of PERK, iNOS, Arg1, and β-actin from PERK wild-type and deficient BMDMs treated with LPS plus IFN-γ (M1), IL-4 (M2), or Thapsigargin. Data are representative of two independent experiments. j-l, Representative histogram (left) and quantitative plot (right) of iNOS (j), TNF-α (k), or expression of CD206 and CD301 (l) in BMDMs stimulated with LPS plus IFN-γ (M1) or IL-4 (M2) (n = 3; mean ± s.e.m). Data are representative of three independent experiments. m, Immunoblot analysis of p-PERK, PERK, XBP1s, and β-actin in unstimulated (M0), LPS + IFN-γ (M1), or IL-4 (M2) stimulated BMDMs. Data are representative of three independent experiments. All data were analyzed using two-tailed unpaired Student’s t-test (e,g,j,j,k,l) or one-way ANOVA with Tukey’s multiple comparisons test (f).
