SRX18307768 GSM6742512_r1 SRP408627 PRJNA903157 SRS15798422 GSM6742512 GSM6742512 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA Sensory epithelium from cochlear explants was dissociated to single cells using 0.25% trypsin-EDTA for 15 min incubation at 37C. Tissues were gently singularized by P1000 pipette in a complete medium (DMEM/F12 medium containing 10% FBS). Cells were strained twice through 40 mm mesh into 5 ml medium, spun down for 5 min at 300 g to pellet, and resuspended in 50 ml of complete medium. Roughly 20000 cells were recovered from both groups after 10X Chip-G running. Single cells from cochlea pretreated with Drug before noise exposure and untreated (noise-exposed-only) cochleae were captured separately to prepare cDNA libraries. NIHL (Noise-only; n=5) vs. Drug-NIHL (n=5). Single-cell suspensions were immediately transferred to individual channels of the chip (Chromium Next GEM Chip G) to generate gel beads in emulsion together with reverse transcription master mix on the droplet-based high-throughput Chromium Controller (10X Genomics) allowing cell lysis, individual cell barcoding, and reverse transcription by Chromium Next GEM Single Cell 3' Kit v3.1 (10X Genomics, PN-1000128). Briefly, single-cell library preparation was carried out by emulsion breakage, PCR amplification, cDNA fragmentation, oligo adapter, and Illumina sample index addition following the manufacturer's protocol. The single-cell derived cDNA libraries were then assessed with a bioanalyzer (High Sensitivity DNA Kit, Agilent 2100 Bioanalyzer) for quality control. Following library preparation, next-generation sequencing was performed with paired-end sequencing of 150 bp each end using Illumina HiSeqXTen (Novogene Inc.). HiSeq X Ten SRX18307769 GSM6742513_r1 SRP408627 PRJNA903157 SRS15798423 GSM6742513 GSM6742513 RNA-Seq TRANSCRIPTOMIC SINGLE CELL cDNA Sensory epithelium from cochlear explants was dissociated to single cells using 0.25% trypsin-EDTA for 15 min incubation at 37C. Tissues were gently singularized by P1000 pipette in a complete medium (DMEM/F12 medium containing 10% FBS). Cells were strained twice through 40 mm mesh into 5 ml medium, spun down for 5 min at 300 g to pellet, and resuspended in 50 ml of complete medium. Roughly 20000 cells were recovered from both groups after 10X Chip-G running. Single cells from cochlea pretreated with Drug before noise exposure and untreated (noise-exposed-only) cochleae were captured separately to prepare cDNA libraries. NIHL (Noise-only; n=5) vs. Drug-NIHL (n=5). Single-cell suspensions were immediately transferred to individual channels of the chip (Chromium Next GEM Chip G) to generate gel beads in emulsion together with reverse transcription master mix on the droplet-based high-throughput Chromium Controller (10X Genomics) allowing cell lysis, individual cell barcoding, and reverse transcription by Chromium Next GEM Single Cell 3' Kit v3.1 (10X Genomics, PN-1000128). Briefly, single-cell library preparation was carried out by emulsion breakage, PCR amplification, cDNA fragmentation, oligo adapter, and Illumina sample index addition following the manufacturer's protocol. The single-cell derived cDNA libraries were then assessed with a bioanalyzer (High Sensitivity DNA Kit, Agilent 2100 Bioanalyzer) for quality control. Following library preparation, next-generation sequencing was performed with paired-end sequencing of 150 bp each end using Illumina HiSeqXTen (Novogene Inc.). HiSeq X Ten