Fig. 4: Adaptive and innate immune cells in control and alectinib-treated EA1 and EA2 tumors.

EA1 or EA2 cells were implanted in the left lung of C57BL/6 mice. Tumors were established for 3 weeks and then mice were treated for 4 days with either control or 20 mg/kg alectinib. a Tissue was harvested, processed, and FFPE slides were submitted to multispectral imaging using Vectra Polaris. Data acquisition, segmentation, and cell phenotyping was performed using inForm software. The R package Akoya Biosciences phenoptrReports was used to count CD3+/CD8+ and CD3+/CD8− cells (presumed to be CD4+ T cells) as well as B220-positive B cells. Differences between groups was analyzed by the Kruskal-Wallis test with correction for multiple comparisons. b After 4 days of treatment, mice were sacrificed and a single-cell suspension was made from the left tumor-bearing lung. The single cell suspension was stained and submitted to flow cytometry analysis for innate immune cells using the previously described gating strategy³⁰. Differences between groups were analyzed with ordinary one-way ANOVA and correction for multiple comparisons. Data are presented as the mean ± SEM where *, **, ***, and **** indicate p values <0.05, <0/01, <0.001, and <0.0001, respectively.