Extended Data Fig. 7: AMBRA1 controls tumour growth in a mouse model of lung cancer.
From: AMBRA1 regulates cyclin D to guard S-phase entry and genomic integrity
a, Schematic representations of the mouse model and initial testing of the system. The KrasG12D transgenic mouse is mated with the conditional Ambra1flox/flox mouse to produce the Ambra1+/+::KrasG12D/+ and the Ambra1flox/flox::KrasG12D/+ genotypes. Lung-specific expression of oncogenic KrasG12D and deletion of Ambra1 is induced by intranasal inoculation with defective adenoviral particles carrying the Cre recombinase. b, Immunoblot analysis of AMBRA1 immunoprecipitation from tissue lung samples from Ambra1flox/flox::KrasG12D/+ mice 16 weeks after administration of AdenoCre (n = 3). c, The expression of the Ambra1 floxed allele after Cre administration was verified by RT–PCR performed in lung tissue samples as in c (n = 3). Primers were designed to distinguish wild-type and floxed alleles. d, Representative examples of H&E images of fixed lungs. Bottom, Magnification of the bronchus, highlighting the tumour initiation site. Scale bar, 1 mm. e, Quantification of immunohistochemistry staining in Fig. 3b (Ki67, n = 3; γH2AX, n = 3; RPA(pS4/8), n = 3; cyclin D1, n = 4; c-Myc(pS62), n = 3 in two independent tumours for each condition). Unless otherwise stated, n refers to biologically independent samples; data are mean ± s.e.m. P values for γH2AX and cyclin D1 by two-tailed Welch t-test; P values for Ki67, c-MYC(pS62), RPA(pS4/8) by two-tailed unpaired t-test.
