Journal: bioRxiv
Article Title: Cancer-associated adipocytes mediate CD8 + T cell dysfunction via FGF21-driven lipolysis
doi: 10.1101/2024.10.18.618991
Figure Lengend Snippet: A. GSEA analysis of gene signatures of mitochondrial dysfunction of CD8 + TILs sorted from MC38 tumors in Fgf21 fl/fl and Fgf21 ΔAD mice (n = 3 mice). B. Transmission Electron Microscope (TEM) analysis of mitochondrial morphology in CD8 + T cells treated with conditioned media from WT-AAD or E0771 tumor-adjacent CAAs from Fgf21 fl/fl and Fgf21 ΔAD mice after activation by anti-CD3/CD28 for 96 hr (n = 4 biological replicates). C. Seahorse measurement of oxygen consumption rate (OCR) in CD8 + T cells treated with conditioned media from WT-AAD or E0771/MC38 tumor-adjacent CAAs from Fgf21 fl/fl and Fgf21 ΔAD mice after activation by anti-CD3/CD28 for 96 hr (n = 3 biological replicates). D. FACS analysis of ROS hi CD8 + T cells treated with conditioned media from WT-AAD or E0771/MC38 tumor-adjacent CAAs from Fgf21 fl/fl and Fgf21 ΔAD mice after activation by anti-CD3/CD28 for 96 hr (n = 3 biological replicates). E. FACS analysis of the percent of PD-1 hi ROS hi of CD8 + T cells treated with conditioned media from WT-AAD or E0771/MC38 tumor-adjacent CAAs from Fgf21 fl/fl and Fgf21 ΔAD mice after activation by anti-CD3/CD28 for 96 hr (n = 3 biological replicates). F. FACS analysis of the percent of PD-1 hi , LAG-3 + , TIM-3 + of CD8 + T cells treated with conditioned media from WT-AAD or E0771/MC38 tumor-adjacent CAAs, or plus with N-Acetylcysteine (NAC), after activation by anti-CD3/CD28 for 96 hr (n = 3 biological replicates). G. FACS analysis of the percent of IFNγ + , Granzyme B + , TNFα + and Perforin + of CD8 + T cells treated with conditioned media from WT-AAD or E0771/MC38 tumor-adjacent CAAs, or plus with NAC, after activation by anti-CD3/CD28 for 96 hr (n = 3 biological replicates). Data are presented as the mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001 by Student’s t test (B, C, D, E) and two-way ANOVA (F, G).
Article Snippet: Cells were fixed with electron microscope fixative buffer (cat#G1102, Servicebio), post-fixed in 1% osmic acid solution, dehydrated with a graded ethanol series, pure acetone and embedded in embedding agent.
Techniques: Transmission Assay, Microscopy, Activation Assay
