H1703

The lung cancer cell lines NCI-H226 and H1703 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences. Cells were cultured with DMEM containing 10% serum and penicillin-streptomycin solution. Cells were placed in an incubator at 37° and 5% CO₂. Cell transfection was mainly performed with Lipofectamine 2000 and was conducted according to the manufacturer’s instructions. Forty-eight hours after cell transfection, 1 µg/mL of puromycin was used for screening. Seven days later, the surviving cells were mixed, and gene expression was detected with Western blot analyses.

Human NSCLC cell lines (A549 and H460) were obtained from the Cell Resource Center, Peking Union Medical College (Beijing, China), H1299 was purchased from the ATCC, and other human NSCLC cell lines (H292, H2228, HCC827, H1703, and Calu-1) were purchased from Shanghai Cell Bank of Chinese Academy of Sciences (Shanghai, China). The murine Lewis lung carcinoma (LLC) cell line was obtained from the ATCC. The LLC-OVA cell line expressing a single polypeptide encoding H-2Kbb2-M, and the OVA SIINFEKL peptide was kindly gifted by Amer A. Beg (Moffitt Cancer Center, Tampa, FL). H460, H1299, H292, H2228, HCC827 and H1703 cell lines were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. Calu-1 cells were cultivated in McCoy’s 5A medium supplemented with 10% FBS and 1% penicillin-streptomycin. A549 and LLC-OVA cells were cultured in DMEM medium supplemented with 10% FBS and 1% penicillin-streptomycin. All the cell lines were cultured in a 37 °C humidified incubator with 5% CO₂.
Anlotinib was kindly provided by Chia Tai Tianqing Pharmaceutical Group Co., Ltd (Nanjing, Jiangsu, China) and was prepared as a 10 mmol/L stock solution in dimethyl sulfoxide (DMSO) for in vitro experiments or in normal saline for in vivo studies.