Fig. 4

Jnk1/2 ablation accelerated Lkb1-deficiency-induced LSCC progression. a Quantification of cell colony formation assay of mLSCC^LP.3 cells after knockout of Jnk1/2 or scramble (Control) using Lentiviral Cas9/gRNA. Error bar is SEM; T test: ***P < 0.001. b Representative H&E and IHC staining of 4-month-old (4M) and 7-month-old (7M) CCSP^iCreLkb1^f/fJnk1^f/fJnk2^−/− (Lkb1^d/dJnk1^d/dJnk2^−/−) mouse lungs. SCC markers: p63 and CK5; AD marker: TTF1. Scale bar: 600 μm (black box, 4 months), 60 μm (black box, 7 months), and 10 μm (solid black). c Representative H&E and IHC staining of 7-month-old Lkb1^d/dJnk1^d/dJnk2^−/− mouse lung epithelial hyperplasia (I), and adenocarcinoma (II). Scale bar: 300 μm (black box, left panel), 60 μm (black box, middle and right panels). d The representative H&E and IHC staining of lung adenosquamous carcinoma in 7-month-old Lkb1^d/dJnk1^d/dJnk2^−/− mice. Adenocarcinoma (AD) cells are indicated by black arrows, and squamous cell carcinoma (SCC) cells in the lower half of the image are indicated by the green arrowhead. A large portion of the AD component has positive staining for the typical SCC markers CK5 or p63 while having negative or weak staining for TTF1. The SCC component has positive staining for CK5 and p63 while having negative staining of TTF1. Scale bar: 200 μm. e WB analysis of protein expressions in mLSCC^LP.3 cells under 5-h treatment of IL-1β (2 ng/mL), TNFα (20 ng/mL) and Anisomycin (10 μM). Free medium and culture medium are described in the Methods section