Fig. 2: Histological and molecular analysis of lung tumors in Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL mice.

A Representative images of lung sections from mice with indicated genotypes sacrificed at the humane endpoint, stained with H&E (Scale bars = 5 mm (top) and 100 µm (bottom)) B Representative images of lung sections from mice with indicated genotypes immunostained for CD56 (Scale bars = 2.5 mm and 5 mm (top) and 50 µm (bottom), n = 8 for Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL, n = 11 for Rb1^FL/FL Tp53^FL/FL) C Representative images of lung sections from mice with indicated genotypes immunostained for Ki67 and graph depicting quantification of Ki67 positive cells. Mann–Whitney test. Mean ± SEM. (Scale bars 2.5 mm (top) and 50 µm (bottom), n = 7 for Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL, n = 13 for Rb1^FL/FL Tp53^FL/FL). D Representative images of lung sections from mice at the humane endpoint immunostained for CD45 and graph showing quantification of infiltrating CD45 positive cells. Mann–Whitney test. Mean ± SEM. (Scale bars = 100 µm (top) and 25 µm (bottom), n = 8 for Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL, n = 13 for Rb1^FL/FL Tp53^FL/FL). E, F Principal component analysis (PCA) E and Volcano plot Fof RNA seq data from tumor tissues of Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL (n = 5) compared to Rb1^FL/FL Tp53^FL/FL (n = 6) mice. Genes that were found significantly upregulated (p < 0.01, log2(|FC | ) > 2) or downregulated (p < 0.01, log2(|FC | ) < 2) in tumors from Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL compared to Rb1^FL/FL Tp53^FL/FL mice are indicated with red or blue dots, respectively. G Representative immunoblot analysis of protein extracts from SCLC cell lines isolated from Rb1^FL/FL Tp53^FL/FL and Rb1^FL/FL Tp53^FL/FL Nemo^FL/FL mice at the humane endpoint (n = 4). Uncropped images of immunoblots are presented in Supplementary Fig. 1.