Extended Data Fig. 8: Dietarily reprogramed TAMs engulf apoptotic cells in MYC-PyMT tumors.
From: Reprogramming tumour-associated macrophages to outcompete cancer cells
a,b, Experimental design, flow cytometric analysis and quantification of TAM engulfment of apoptotic cells in a complete or low amino acid (AA) RPMI medium (1: 4 mixture of complete RPMI and AA-free RPMI). Apoptotic PyMT cancer cells were labeled by CellTrace Violet (CTV) and induced by H2O2 treatment. TAMs were isolated from tumors of control-PyMT and MYC-PyMT mice (n=5 for each group) treated with a low protein (LP) diet (2% protein in weight, 5BT9, TestDiet). c, Immunofluorescence images showing expression or localization of F4/80, EpCAM, and TUNEL in 6 mm x 6 mm tumors from control-PyMT and MYC-PyMT mice under LP or normal protein (NP) diet (15% protein in weight, 5CC7, TestDiet) conditions. Scale bar: 50 μm. Representative macrophages marked by yellow dashed lines. d, Quantification of percentage of TUNEL-positive cells among all cells. e, Quantification of percentage of TUNEL-positive nuclei inside F4/80-expressing macrophages. Data points are average percentage from random captured images. n = 3 mice for each group in (d,e). f, A schematic diagram showing the generation of bone marrow (BM) chimera mice. BM cells from Fcgr1-cre-Rosa26LSL-H2B-mCherry/+ mice were transferred into lethally irradiated MYC-PyMT recipients with a tumor burden around 250-350 mm3 followed by switching to the LP diet two weeks after. g, Immunofluorescence images showing expression or localization of F4/80, EpCAM, H2B-mCherry, and TUNEL in 6 mm x 6 mm tumor tissue samples from BM chimera mice described in (f). Scale bar: 10 μm. All statistical data are shown as mean ± S.D. One-way ANOVA with the Tukey multiple comparison test correction in (b,d,e).
