library_construction_protocol: 3 days after ligature placement, the gingival tissue was harvested and incubated with RPMI 1640 medium containing 0.15 mg/ml DNase and 3.2 mg/ml type IV collagenase for 25 minutes at 37°C in a shaker. 0.5M EDTA (pH8) was added to the suspension to stop the enzymatic digestion. After homogenization, the dissociated suspension was passed through a 70mm cell strainer, washed, and incubated with an anti-mouse CD45 antibody-Alexa Fluor 700 for half an hour. After washing, Hoechst was added to the cell suspension. Then, live CD45+ cells were immediately sorted by a Cytek Aurora CS cells sorter and counted under a microscope. Samples with a viability above 85% and more than 990 live cells/µl were proceeded with scRAN-seq. Library was performed according to the manufacturer's instructions (Chromium Next GEM Single Cell 3' Reagent Kits v3.1 protocol, 10X genomics) Briefly, sorted CD45+ gingival cells were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was reverse-transcribed to cDNA, which contains an Illumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10X Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified with PCR and the appropriate sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction, Illumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.