GSM7527830: YUMMER1.7 (YR) tumor (8 days post-tumor initiation, untreated), scTCR-seq; Mus musculus; OTHER
24 hours post-treatment (if applicable), untreated and TTx-treated tumors for scRNA-seq were minced in RPMI with 2% FBS, then incubated with agitation at 37C, with 1 mg/mL Collagenase IV (C5138, Sigma-Aldrich) and 0.1 mg/mL DNAse I (10104159001, Roche) for 30 minutes. Tumors were then filtered through 70 µM filters into a single-cell suspension. Samples were then FACS-sorted into three populations: CD45+CD3+ (T cells), CD45+CD3- (non-T immune cells), CD45-CD3- (tumor and stroma). The populations were then recombined in a 2:1:1 ratio, and 10,000 cells from each condition (TTx-treated and control) were loaded onto the 10X Chromium System for sequencing. Antibodies used for sorting: CD45 (I3/3.2), CD3 (17A2) VDJ library construction was performed according to the manufacter's instructions (single cell 5' v1 protocol, 10x Genomics). scTCR-seq