Extended Data Fig. 6: Macrophage-specific ablation of LPCAT3 abolishes GW3965 anti-tumor responses.
a, b, qPCR analysis of mRNA expression of sXBP1 (a) and immunoblot and quantification (b) of indicated proteins in WT or LPCAT3-KO BMDMs exposed to regular culture medium (Ctrl) and YUMM1.7 CM in absence or presence of 3μM GW3965 (n = 9 per group for qPCR and n = 3 per group for immunoblots). Data are pooled from three independent experiments c, Proliferation of CFSE-labeled T cells activated with anti-CD3 and anti-CD28 alone or co-cultured with WT or LPCAT3-KO BMDMs previously treated with YUMM1.7 CM in the absence or presence of GW3965 in a ratio 2:1 for 72 h (n = 3 per group). Data are representative of three independent experiments. d, Illustration of experimental design for bone marrow transplantation. e, qPCR analysis of exon 3 of LPCAT3 gene in LPCAT3fl/fl (WT) and LysM-Cre LPCAT3fl/fl (KO) mice (n = 15). f, Bone marrow was isolated from WT and KO chimeric tumor-bearing mice and the abundance of indicated immune cells was measured by flow cytometry (n = 4). g, Percentage of mTAMs among CD11b+ tumor-infiltrating myeloid cells from YUMM1.7-OVA melanoma treated with either control vehicle or GW3965 in mice transplanted with BM cells from LPCAT3fl/fl (WT) and LysM-Cre LPCAT3fl/fl (KO) mice (WT + Vehicle: n = 9; WT + GW3965: n = 10; KO + Vehicle: n = 10; KO + GW3965: n = 9). Data are pooled from two independent experiments. Each symbol represents one individual. All data are mean ± s.e.m and were analyzed by two-tailed, unpaired Student’s t-test (a, e-g), RM one-way ANOVA with Bonferroni’s multiple comparison test (b), and ordinary one-way ANOVA with Tukey’s multiple comparison test (c).
