Pd 1 antibody

C57BL/6 mice (5 weeks old, female) were purchased from the Experimental Animal Center of Three Gorges University. After one week of adaptive feeding, 1*10^6 B16-F10 melanoma cells were injected subcutaneously. When the diameter of the tumor was 5–9 mm, the mice were randomly divided into four groups. There were six mice in each group. The control group was given 100 μl PBS, the cromolyn sodium group was given 100 μl 2 mg/ml cromolyn sodium (Sigma, St Louis, MO, USA) every day, the PD-1 antibody group was given 100 μl 2 mg/ml PD-1 antibody (BIOXCELL, New Hampshire, USA) once every 3 days, and the combined group was given 100 μl 2 mg/ml PD-1 antibody once every 3 days and 100 μl 2 mg/ml cromolyn sodium every day. cromolyn sodium was dissolved in normal saline and administered by intraperitoneal injection [11 (link)]. The PD-1 antibody was diluted with a special diluent produced by BIOXCELL and administered by intraperitoneal injection. The long diameter (L) and short diameter (W) of the tumors were measured every two days, and the volume was calculated according to the formula: V = LW²/2. The tumors were collected after 15 days.

Mice were provided by Hunan Westlake Jingda Laboratory Animal Co., Ltd. (Changsha, China), and the animal experimental protocol was approved by the Animal Ethics Committee of Huazhong University of Science and Technology and conducted accordingly (S3563). Mice were randomly assigned to the indicated groups. Tumor volumes were calculated according to the formula (L × W2)/2. HCT116 cells were infected with shControl, shFH, shPCSK9, or shFH+PCSK9 lentivirus after stable screening. Nude mice were subcutaneously inoculated with 100 μL of HCT116 cell suspension corresponding to approximately 2 × 10⁶ cells to establish a subcutaneous tumor model. Mouse MC38 cell lines infected with shControl and shFh1 lentiviruses were stably screened. C57BL/6 mice were subcutaneously inoculated with 100 μL of MC38 cell suspension to establish a C57BL/6 mouse subcutaneous tumor model. PD-1 antibody (Bioxcell, Cat No. BP0146, Lebanon, NH, USA) and isotype IgG antibody (Bioxcell, Cat No. BE0089, Lebanon, NH, USA) against MC38 subcutaneous tumor models were administered by intraperitoneal injection at a dose of 5 mg/kg every 3 days. A PCSK9 inhibitor (Selleck, Cat No. 489415-96-5, Houston, TX, USA) and DMSO were administered subcutaneously at a dose of 4 mg/kg/day.

Xenograft experiments were performed using C57BL/6 mice to assess the role of circNCOA3 in influencing the efficacy of PD-1 antibody treatment. Briefly, a total of 1 × 10⁶ cells (MC38-sh-circNCOA3 or MC38-sh-NC) were implanted subcutaneously in the left flank of 6-week-old C57BL/6 mice. Each mouse population was randomly divided into two groups and injected intraperitoneally with either PBS or PD-1 antibody (Bio X Cell, West Lebanon, NH, USA) according to a previously described method^{[26 (link)]}.