Fig. 3: Restraint stress significantly decreases T-cell infiltration into 66CL4 mammary tumours but does not affect the primary tumour weight and volume.
From: An integrated framework for quantifying immune-tumour interactions in a 3D co-culture model
Female BALB/c mice were injected with 66CL4 cells into the fourth mammary fatpad. Two weeks post injection mice were randomised into stress (n = 6 biologically independent animals) and nonstress (n = 6 biologically independent animals) groups. Mice from the stress group were subjected to 2 h of daily restraint stress for two weeks. Mammary tumours were collected and paraffin-embedded for immunohistochemical analysis. a Tumour sections were fluorescently labelled with the T-cell marker CD3ε (Cell Signalling, #99940) and nuclear stain DAPI (Sigma, D9542). Analysis of CD3 expression in the mammary tumours of stressed and non-stressed mice was done using CellProfiler, where the percentage of CD3-positive cells was calculated compared to the total number of cells (%CD3+/total cells). The scale bar represents 40 µm. The amount of immune cell–cell infiltration inside the mammary tumours of stressed mice is significantly reduced compared to the non-stressed mice. b No significant difference in primary tumour volume between stressed and nonstressed mice. c No significant difference in primary tumour weight between stressed and non-stressed Statistical significance was calculated using a two-tailed Mann–Whitney test, data represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.
