Quantification of how combination serine/threonine-protein kinase B-Raf inhibitor (BRAFi) and anti–programmed death ligand 1 (aPDL1) affects tumor-associated macrophage (TAM) accumulation and nanomedicine delivery. A, Representative confocal microscopy of optically cleared tumors reveals macrin accumulation at 24 hours after administration, following an 8-day course of treatment with combination BRAFi and aPDL1 in the murine TBP model of anaplastic thyroid cancer (ATC; scale bar, 5 mm). B, Corresponding to A, the scatter dot plot shows TAM density quantified across single cells from microscopy data by using macrin to identify TAM. Bonferroni correction applied for multiple hypothesis testing (n = 2 hypotheses, corresponding to lung and thyroid, comparing with or without combination BRAFi and aPDL1 treatment). C, Graph shows macrin and drug delivery nanoparticle (DDNP) uptake correlated across treatment groups on a tumor-by-tumor basis (see Fig E11 [online] for individual tumor-by-tumor correlation within individual subjects). D, Scatter dot plots show DDNP uptake quantified by confocal microscopy in TAM (macrin+) and other (macrin-) cells by using the TBP model as a fraction of average uptake in macrin+ cells. See Figure E9 (online) for comparison across drug treatments. For B and D, data are means ± standard deviation.