library_construction_protocol: All the tumors were extracted 20 days after tumor implanted. Total RNA was extracted from GAD1-shA GAD1-shB and NTC MC38 tumor samples (3 replicates per group) using an RNeasy kit (Qiagen, Cat# 74104) and dissolved in RNase-free water. RNA samples were sequenced on Illumina novaseq. Sequencing libraries were generated using NEBNext® UltraTM RNALibrary Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations andindex codes were added to attribute sequences to each sample.