Ab192579

For western blotting, equal amounts of proteins were resolved by 10% SDS-PAGE (CW Biotech Co., Ltd.). Proteins were blotted onto a methanol-charged PVDF membrane using a wet transfer system (70 V, 2 h). The membrane was blocked with 5% bovine serum albumin for 1.5 h at room temperature, washed with Tris-buffer saline containing 0.1% Tween-20 (TBST) three times (5 min each time), and then incubated with the primary antibodies anti-PXR (1:1000, ab192579, Abcam, Cambridge, UK), anti-CYP3A2 (1:1000, ab195627, Abcam), or anti-GAPDH (1:4000, ab94282, Abcam). The membrane was washed with TBST five times (5 min each) and incubated for 1 h with goat anti-rabbit IgG H&L (HRP) secondary antibody (1:5000, ab6721, Abcam). After five washes with TBST, the protein bands were detected by chemiluminescence reagents (ECL kit ab133406, Abcam). An imaging system (ImageQuant LAS 500, GE Heathcare Life Sciences, Chicago, IL, USA) was used to visualize the protein bands, and densitometry was performed with Image J software. The density of each immunoreactive band was normalized to the density of its corresponding GAPDH band.

Raw PDT leaves, FBT, and Hua Juan Tea (HJT) were purchased from the Baishaxi Tea Factory Co., Ltd. (Yiyang, Hunan, China). The Eurotium cristatum strains were isolated from Baishaxi brick tea using the direct separation technique. Tea-lyophilized powder was prepared at the Hunan Agricultural University. Cigarettes were purchased from the Hunan China Tobacco Industry Co., Ltd. (Changsha, Hunan, China). Vacuum diaphragm pumps were purchased from Kamoer, KVP15-KL-1 (Shanghai, China). Analytical kits for GSH-Px, SOD, and MDA were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China). ELISA kits for the analysis of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), IL-8, and IL-1β were purchased from Wuhan Hualianke Biotechnology Co. Ltd. (Wuhan, Hubei, China). Antibodies against p-p38 (4511S), p38 (8690S), p-Jun N-terminal kinase (p-JNK, (9251S)), JNK (9252S), p-extracellular-regulated kinase (p-ERK, (4370S)), and ERK (4695S) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Monoclonal antibodies against PXR (ab192579), AhR (ab84833), and GAPDH (ab181602) were purchased from Abcam (Cambridge, UK). All other chemicals and reagents were at analytical grade.

Analytical kits for SOD, GSH-Px and MDA were purchased from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). ELISA kits for the analysis of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), IL-8, and IL-1β were purchased from Wuhan Hualianke Biotechnology Co. Ltd. (Wuhan, China). Extracellular signal-regulated kinases (ERK), phospho-ERK (p-ERK), c-Jun N-terminal kinase (JNK), phosphor-JNK (p-JNK), p38 and phosphor-p38 (p-p38) were purchased from Cell Signaling Technology (Danvers, MA, USA). Monoclonal antibodies against PXR (ab192579), AhR (ab84833), and GAPDH (ab181602) were purchased from Abcam (Cambridge, UK). All other chemicals and reagents were of analytical grade.