Recombinant human tgf β1 tgf β

The human lung adenocarcinoma cell lines A549, PC9 (EGFR mutant) and the human colon cancer cell lines SW480 and SKCO1 (KRAS mutant) were obtained from the China Center for Type Culture Collection (Wuhan, China). A Cisplatin-resistant subline of A549 and a gefitinib-resistant subline of PC9 were established by repeated subculturing with gradual increasing in the Cisplatin (2, 4, 6, 8 and 10 μM) or gefitinib concentration (4, 8, 16, 20 and 40 μM) over a 3-month period.^{52 (link)} In addition, A549, PC9, SW480 and SKCO1 cell lines with stable overexpression or knockdown of ZNF32 were constructed as described previously.^{35 (link)}BALB/c male nude mice, 6 weeks of age, were maintained under standard conditions in the animal facility of Sichuan University. All experiments in this study were performed in accordance with the nation's relevant laws and animal welfare requirements.
Cisplatin and 5-FU were purchased from Sigma-Aldrich (St Louis, MO, USA), and gefitinib and AZD6244 were obtained from AstraZeneca (Cheshire, UK) and dissolved in DMSO to obtain a stock solution of 10 mM. These drugs were diluted with culture medium for use in the experiments. Recombinant human TGF-β1 (TGF-β) was purchased from R&D Systems and used at a concentration of 10 ng/ml. LY2157299 (Selleck), an inhibitor of TGF-β signaling, was used at a concentration of 1 μM in the experiments.

Three TNBC cell lines (MDA-MB-231, Hs578T, and MDA-MB-157) were purchased from the American Type Cell Collection (Manassas, VA), and Mx-1 cells were purchased from CLS Cell Lines Service (Eppelheim, Germany) in 2016, and passaged in our laboratory for less than 6 months after receipt or resuscitation. All cell lines were tested monthly for mycoplasma contamination using the MycoAlert mycoplasma detection kit (Lonza Walkersville, Inc, Walkersville, MD) and were cultured for no more than 20 passages from the validated stocks. All cell lines were cultured in RPMI with 10% FBS at 37.0° C and 5% CO₂. Eribulin-resistant MDA-MB-231 and Hs578T cells were previously established in our laboratory [23 (link)].
Eribulin was purchased from Eisai Co., Ltd. (Tokyo, Japan). Paclitaxel for in vitro use was purchased from Sigma-Aldrich (Saint Louis, MO) and for in vivo use was purchased from Nippon Kayaku Co., Ltd. (Tokyo, Japan). Recombinant human TGF-β1 (TGF-β) was purchased from R&D Systems (Minneapolis, MN). TGF-β receptor I kinase inhibitor (LY2157299) was purchased from Sellek Chemicals (Houston, TX).