Ab68515

Lung tissues were separated, fixed, dehydrated, embedded and cut into slices (5 µm) in sequence. After being dewaxed and hydrated, slices were stained with Wiegert's solution for 10 min and differentiated with acidic ethanol for 10 s. Subsequently, slices were treated with Masson bluing buffer for 5 min, ponceau-fuchsin solution for 10 min, phosphomolybdic acid solution for 3 min, aniline blue solution for 5 min, and weak acid solution for 30 s in turn. Finally, slices were dehydrated with 95% ethanol and absolute ethanol, transparentized with dimethylbenzene, mounted with neutral resin, and imaged under a digital trinocular camera microscope (CX23, Olympus). transferred onto a PVDF membrane (EMD Millipore, Billerica, MA, USA). After being blocked with 3% bovine serum albumin (BSA, Solarbio) at room temperature for 1 h, membranes were hatched with primary antibodies against diverse proteins, containing p38 (1:1000, ab68515), phosphorylated p38 (p-p38, 1:2000, ab196495), β-catenin (1:1000, ab16051), c-Jun N-terminal kinase (JNK, 1:1000, ab32072), p-JNK (1:1000, ab32072) and GAPDH (1:2500, ab9485; all from Abcam) at 4°C overnight. Membranes were then treated with corresponding secondary antibodies at room temperature for 3 h and visualized by an ECL assay (Beyotime). The gray value was calculated by QUANTITY ONE software.