Stat3 p stat3tyr705

Nifuroxazide was purchased from (Xiyashiji Chemical Co., LTD, ChengDu, Sichuan, China). Purity (98%) was measured by high-performance liquid chromatograph analysis. For in vitro assays, nifuroxazide was prepared initially as a 20 mM stock solution in dimethyl sulfoxide (DMSO) and stored at −20 °C. Then the stock solution diluted in the relevant assay medium, and 0.1% DMSO served as a vehicle control. For in vivo studies, nifuroxazide was prepared in 40% (v/v) polyethylene glycol 400 containing 5% (v/v) propylene glycol and dosed at 0.1 ml/10 g of body weight.
MTT, DMSO and Hoechst 33258 were from Sigma Chemical Co. (St Louis, MO, USA). The primary antibodies against Stat3/P-Stat3^Tyr705, MMP-2, -9 (MMP-2, MMP-9), CC-3, Bcl-2 and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). FITC-CD11b, PE-Gr1 conjugated antibodies were obtained from BD Biosciences (San Diego, CA, USA). Mouse monoclonal anti Ki-67 was purchased from Merck-Millipore (Bill, MA, USA). The Annexin V-FITC Apoptosis Detection Kit was purchased from KeyGen Biotech (Nan-jing, China).

All reagents, unless otherwise noted, were purchased from Sigma Chemical Co. (St Louis, MO, USA). Hoechst 33258 and the Annexin V-FITC Apoptosis Detection Kit were purchased from KeyGen Biotech (Nanjing, China). And 0.5% crystal violet was from Beyotime (Beijing, China). The primary antibodies against Stat3/p-Stat3^Tyr705, MMP-9, cleaved caspase-3, Ki-67, Bax, and Bcl-2 were obtained from Cell Signaling Technology (Beverly, MA, USA). β-Actin and MMP-2 were purchased from ZSJQ-BIO Co. (Beijing, China) and Merck Millipore (Billerica, MA, USA), respectively. FITC-CD8a-, FITC-CD4a-, and PE-CD69-conjugated antibodies were obtained from BD Biosciences (San Diego, CA, USA).
Pec. (PubChem CID: 5320438) was purchased from Weikeqi Biological Technology Co., Ltd. (Chengdu, Sichuan, China) and has the chemical structure shown in Supplemental Figure 1. The purity was no less than 98% as determined by HPLC, according to the documentation from the manufacturer. For in vitro studies, Pec. was dissolved in DMSO at a stock concentration of 40 mM and stored at −20°C away from light. The fresh solution was prepared every 2 weeks. And it was diluted in relevant cultured medium at a final DMSO concentration of 0.1% (v/v). For in vivo experiments, Pec. was diluted in 5% DMSO, 35% PEG-400, and 60% physiological saline solution.

Nifuroxazide was purchased from Xiyashiji Chemical Co. Ltd (Chengdu, Sichuan, China), and was measured by ¹H-NMR, ¹³C-NMR and ESI-MS analysis. For the in vitro assays, Nifuroxazide was prepared in dimethyl sulfoxide (DMSO) at a stock concentration of 20 mM and stored at −20 °C. Then, it was diluted in the relevant medium at a final DMSO concentration of 0.1% (v/v), and the medium with 0.1% DMSO served as a vehicle control. For in vivo studies, Nifuroxazide was dissolved in 25% (v/v) Cremophor EL/ethanol (50 : 50, Sigma Cremophor EL, 100% ethyl alcohol) and 75% ultrapure water.
DMSO, MTT, DCFH-DA, Rh123 and Cremophor EL were from Sigma Chemical Co. (St. Louis, MO, USA). Hoechst 33258 and NAC were obtained from Beyotime (Beijing, China). The primary antibodies against Stat3/p-Stat3^Tyr705, MMP-2, MMP-9, cleaved caspase-3, Bax, Bcl-2 and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). FITC-CD11b-, PE-Gr1-, FITC-CD8a-, PE-CD69-, PE-CD206- and APC-F4/80-conjugated antibodies were obtained from BD Biosciences (San Diego, CA, USA). Anti-Ki-67 mouse monoclonal was purchased from Merck Millipore (Billerica, MA, USA). The Annexin V-FITC Apoptosis Detection Kit was obtained from KeyGen Biotech (Nanjing, China).