Immunostaining of immune cells from B16F0 tumour sections was performed as described previously (Reynolds et al., 2010 (link)).
Primary mouse lung fibroblasts and endothelial cells were fixed with 4% PFA for 10 min, washed twice and blocked with 5% NGS in PBS for 30 min at room temperature. Primary antibodies to vimentin (1:100; 5741, Cell Signaling), NG2 (1:100; MAB 5385, Millipore), endomucin (1:100; V7C7, Santa Cruz Biotechnology) and α-Sma (clone 1A4, Sigma) were incubated for 1 h at room temperature, washed three times with PBS, and incubated with the relevant secondary antibody for 45 min at room temperature.
Primary mouse lung fibroblasts and endothelial cells were fixed with 4% PFA for 10 min, washed twice and blocked with 5% NGS in PBS for 30 min at room temperature. Primary antibodies to vimentin (1:100; 5741, Cell Signaling), NG2 (1:100; MAB 5385, Millipore), endomucin (1:100; V7C7, Santa Cruz Biotechnology) and α-Sma (clone 1A4, Sigma) were incubated for 1 h at room temperature, washed three times with PBS, and incubated with the relevant secondary antibody for 45 min at room temperature.
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