Novaseq sp 100 v1 kit

Libraries preparations for single-cell immune profiling, sequencing, and post-processing of the raw data were performed at the GIGA-Genomics Core Facility (Belgium). Sorted cells were washed with PBS (calcium and magnesium free) containing BSA (400 μg/mL). 12,800 cells were loaded on Chromium Controller (10x Genomics). Samples were further processed for dropletbased RNA sequencing and libraries were prepared using Chromium Single Cell 3' Reagent Kits v3 (10x Genomics). Amplified cDNA quality controls were performed with an Agilent bioanalyzer (Agilent) and final library profile were checked on Qiaxcel (Qiagen). Sequencing libraries were loaded an Illumina Novaseq sequencer with NovaSeq SP 100 v1 kit (Illumina, CA, USA) using the following read lengths: 28 bp for Read1 (18 bp Barcode + 10 bp Randomer), 8 bp for Sample Index and 88 bp for Read2. Library quantification was processed with KAPA Library quantification kit (KAPA Biosystems). Analysis of scRNA-seq samples is described in supplementary Material and Methods.
Quantification and statistical analysis. For comparisons between two groups, Student's two-tailed t test was used. For comparisons between multiple groups, one-way or two-way ANOVA was used with multiple-comparison tests. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.
Error bars represent SEM.