Figure 2. Temporal dissociation of BRAF^V600E expression from additional genetic events.

A. Lung infection schematic of Braf^FA;Trp53^lox/lox; Rlz (BTR) mice to test the efficiency of Ad-Cre mediated recombination in established BRAF^V600E-driven tumors initiated with Ad-Flp. Below are H&Es stained with β-galactosidase.

B. (i) Schematic of infection (ii) Comparison of temporal separation of TP53 silencing and BRAF^V600E activation to BRAF^V600E alone using two infection strategies. 1^st row, photographs of the macro lung. 2^nd–4^th rows, paraffin lung sections of Braf^FA, BT and Braf^FA; Trp53^Rev (BT^Rev) stained with H&E, Ki67, pERK, and p19^ARF to compare proliferation and MAPK signaling.

C. Paraffin sections of a kidney metastasis detected in a BT^Rev mouse stained with H&E, SPC, NKX2.1, CC10 and AQP5 by IHC and IF.

D. Dot plot showing the average number of lung adenocarcinomas in BT or BT^Rev mice in which BRAF^V600E was activated either prior to or after silencing of TP53.

E. Immunoblot analysis of BRAF^V600E or BRAF^V600E/TP53^Null lung tumor lysates as indicated.

F. Sections of lung tumors from Braf^FA or BT mice were stained to detect BrdU incorporation or for expression of TP53, p21^CIP1 or p27^KIP1 as indicated.