title: GSM7628233: KSQ_001_tumor_1_2_vdj; Mus musculus; RNA-Seq
library_construction_protocol: For the OT1 / B16-Ova model, female C56BL/6J mice were inoculated with 0.5x106 B16-Ova cells subcutaneously in the right flank.    After eight days for small tumor studies (~100mm3 – range of 80mm3-120mm3) or 14 days for large tumor studies (~300mm3 – range of 135mm3-550mm3) mice were randomized by tumor volume and 3x106 sgOlf, sgPD-1 or sgSocs1 OT-1 T cells were transferred in 0.2mL PBS via the lateral tail vein.  Prior to transfer into mice, OT-1s are tested via PCR for a comprehensive list of mouse pathogens by Charles River Laboratory Testing Management.  Tumor volumes were measured twice weekly using a digital caliper and tumor volume (mm3) calculated using the formula: (width2 x length)/2, where length was the longer dimension.  To assess tumor growth following re-challenge in Figure 2, mice undergoing complete tumor rejection were re-challenged with 0.5x106 B16-Ova cells subcutaneously on the left flank on Day 61 and monitored for tumor growth. For the PMEL / MC38-gp100 model: Female C56BL/6J mice were inoculated with 1x106 MC38-gp100 cells.  At the indicated tumor size, mice were randomized and received 7x106 sgOlf, sgPD-1 or sgSocs1 PMEL T cells in 0.2mL PBS intravenously via the lateral tail vein.  For the NOG / hIL-2 NOG engraftment model, female NOG or hIL-2 NOG mice were dosed with 10x106 TIL or KSQ-001 TIL intravenously in 100ml volume. For the TIL / A375-mOKT3 model, female hIL-2 NOG mice were inoculated with 5x106 A375-mOKT3 cells subcutaneously in a 50:50 mix with Matrigel (Corning Biosciences). 7 days post-inoculation, 10x106 TIL or KSQ-001 were transferred intravenously. Droplet-base 5' single-cell RNA sequencing (scRNA-Seq) was performed by the 10x Genomics platform and libraries were prepared by the Chromium Single Cell 5' Reagent kit according to the manufacturer's protocol (10x Genomics, CA, USA).