Hamster igg isotype control

Lymphoma cells expressing MHC class-I (RMA) were labeled with a high CFSE concentration (6 μM), whereas those with defective expression of MHC class-I (RMAS) were labeled with a low CFSE concentration (0.3 μM). The cells were mixed in a 1:1 ratio (1 × 10⁶ cells per cell type) and injected i.p. into C57BL/6J mice for 12 h. To deplete the NK cells, i.p. injections of rabbit anti-NK1.1 (200 μl) or hamster IgG isotype control (Bio X Cell, Cat. Number: 2A3) were performed before injection of CFSE-stained RMA and RMAS cells. Rejection of NK cell-sensitive RMAS relative to NK cell-resistant RMA cells in the peritoneal cavity was calculated as follows: ([CFSE high] control - [CFSE high] experimental group) × 100%.

C57BL/6J mice were inoculated subcutaneously on their right flank with 2 × 10⁶ Lewis lung cells on day 1, and the mice were subsequently randomly separated into groups. After mice developed palpable tumors, mice were i.g. injected with CF (50 mg/kg) or vehicle-injected daily and started on either 0.2 mg anti-PD1 (Bio X cell, Cat. Number: RPM1-14) or hamster IgG isotype control (Bio X cell, Cat. Number: 2A3) every three days intraperitoneally. The mice were sacrificed on day 19 after implantation, and then tumors were excised, weighed, and photographed. The spleen was isolated to prepare splenocytes. Tumor growth was measured by caliper measurements, tumor volume was calculated using the following formula: V = 1/2 × a × b², where V = tumor volume, a = maximum tumor diameter, and b= minimum tumor diameter.