cells were harvested from cell culture plates, washed 3 times with PBS 0.1%BSA Cells were immediately processed using the 10x Genomics Chromium Single Cell 3' Reagent kit v3.1 per manufacturer's instructions. 20,000 cells were loaded for each sample. Cells were partitioned into gel beads, lysed, and barcoded through reverse transcription. cDNA was purified and amplified using appropriate cycle number following the manufacturer's protocol. Library quantification and quality control were performed using High sensitivity DNA ScreenTape kit(Agilent Technologies). Libraries were uniquely indexed using the Chromium dual Index Kit, pooled, and sequenced on an Illumina NovaSeq 6000 sequencer in a paired-end, dual indexing run (28:10:10:90)with a target of 20,000 mean reads per cell.