Fig. 2

Arginase 1 gene and protein expression in the whole tumour and single cells. This figure shows that arginase mRNA and protein were expressed at much higher levels in metastatic than non-metastatic tumours. A Arg1 mRNA levels in 67NR (N = 5) and 66cl4 (N = 5) tumours were measured using qRT-PCR. B Immunoblots of the ARG1 protein and ERK1/2 (loading control) in 67NR (N = 6) and 66cl4 (N = 6) tumours. C–F Representative images of in situ hybridization (C, D) and immunohistochemistry (E, F) showing the spatial distribution of Arg1 mRNA and ARG1 protein in 67NR and 66cl4 tumours. The panels show representative images at 100X magnification, and the inset shows images at 400X magnification. G–I Arg1 mRNA and ARG1 protein levels were significantly higher in CD45⁺ immune cells than in CD45⁻ non-immune cells from 66cl4 tumours than in CD45⁺ and CD45⁻ cells from 67NR tumours. G Arg1 mRNA level in CD45⁺ and CD45⁻ from 67NR (N = 4) and 66cl4 (N = 4) tumours. H Representative flow cytometry plots showing CD45 vs ARG1 protein in single cells from 66cl4 and 67NR tumours. I Percentage of ARG1 positive CD45⁺ and CD45⁻ cells in 67NR (N = 7) and 66cl4 (N = 8) tumours. Values are presented as mean ± SEM and each data point represents a single animal. Statistical significance was determined using the Mann–Whitney test (*p < 0.05, **p < 0.005)