Extended Data Fig. 3: TAMs from mTfeb-/- mice exhibit improved antigen cross-presentation with minimal phenotypic changes.
From: A lysosome-targeted DNA nanodevice selectively targets macrophages to attenuate tumours
a-c, TAMs were isolated from E0771 tumors. a, Quantification of lysosomes in fl/fl and mTfeb-/- TAMs based on LAMP1 immunostaining. Schematic for quantification (left). Quantification of average LAMP1 signal/cell area (n = 10/group) with an average of >40 cells/field (middle). Representative images (right). LAMP1 (red) and DAPI (blue). b, Quantification of lysosomal pH in fl/fl and mTfeb-/- TAMs based on lysotracker staining. Representative flow cytometry image (left). Quantification of relative MFI of lysotracker signal (right). n = 3/group. c, Autophagy gene expression in fl/fl and mTfeb-/- TAMs (left, n = 5). LC3B and p62 protein levels in fl/fl and mTfeb-/- TAMs following treatment with vehicle (Veh) or chloroquine (CQ, 50μM) for 24 h (right). Veh = H2O. Experiment was performed once with n = 3/group. d, M1- and M2-associated gene expression in TAMs from fl/fl and mTfeb-/- E0771 tumors (left, n = 5/group), LLC1 tumors (middle, n = 5/group) and B16F10 tumors (right, n = 4 group). e-f, Quantification of pMel-CD8+ T cell activation (e) and proliferation (f) following co-culture with TAMs isolated from fl/fl and mTfeb-/- B16.OVA tumors. n = 6/group Statistical significance was calculated via two-tailed Student’s t-test (P < 0.05 values are provided); error bars indicate the mean of independent experiments ± s.e.m. ns; not significant. All measurements (n) are biological replicates.
