Figure 7.

Impact of ICI treatment on murine haematopoiesis.

Notes: (a) Pappenheim staining of bone marrow cells in tumor-bearingMsh2^{loxP/loxPVillCre} mice. From each slide, 200 cells were counted and differentiated according to a pre-designed differentiation scheme into the following cell types: myeloblasts, neutrophil pro-/myelocytes, metamyelocytes/neutrophil, eosinophilic pro-/myelocytes, metamyelocytes/eosinophils, lymphocytes, plasma cells, monoblasts, and monocytes. Upper part: representative images of each treatment. Magnification x1000; lower part: Quantification was done-blinded by an experienced researcher. isotype: n = 7 mice; anti-PD1 and anti-LAG-3: n = 6 mice/group. Mean + SD, # p<0.05; ## p<0.01; ### p<0.001; One-way ANOVA (Sidak’s multiple comparisons test’s multiple comparisons test). (b) Bone marrow differentiation using multi-color spectral flow cytometry. A 20-marker panel was applied as described in material & methods. Quantitative analysis of lin⁻c-Kit⁺ precursors. Results show data from 100,000 cells/sample. Mean + SD, each symbol represents one case. Isotype n = 7; $\alpha$-PD1n = 5; $\alpha$-LAG-3 n = 5; # p < 0.05; ## p < 0.01; $ p < 0.05; two-way ANOVA (Tukey’s multiple comparison test). (c) Colony formation assay. The number of viable colony-forming cells was counted after 14 days of in vitro culture. Mean + SD.