NextSeq 500 sequencing; RNA-seq of human colonic cancer cell line CACO-2 transfected 48h with siRNA targeting JAK1 (HJ1D8; HJ1D2) or JAK3 (HJ3D41; HMJ3D1) at 100pM final concentration using LipofectamineRNAiMAX or treated 48h with Tofacitinib (final concentration: 1µM) or Filgotinib (final concentration: 5µM) against controls (transfection medium: Opti-MEM, transfection mixt alone: Opti-MEM + Lipof., siMock, DMSO equivalent to volume added for Filgo 5µM).
RNA-seq of human colonic cancer cell line CACO-2 transfected 48h with siRNA targeting JAK1 (HJ1D8; HJ1D2) or JAK3 (HJ3D41; HMJ3D1) at 100pM final concentration using LipofectamineRNAiMAX or treated 48h with Tofacitinib (final concentration: 1µM) or Filgotinib (final concentration: 5µM) against controls (transfection medium: Opti-MEM, transfection mixt alone: Opti-MEM + Lipof., siMock, DMSO equivalent to volume added for Filgo 5µM).
1 million of CACO-2 cells were seeded in 10 cm Petri dish, then treated or not (see \"treatment protocol\") for 48h before being harvested using trypsine treatment. Cells were washed twice in PBS before RNA extraction. control, cells treated with 0.16% DMSO RNA was extracted using RNeasy Plus Mini Kits (Qiagen cat #74136/74134).  Libraries were generated from 1µg of total RNA using Truseq Stranded mRNA kit (Illumina). Libraries were then quantified with KAPA library quantification kit (Kapa biosystems) and pooled. 4nM of this pool were loaded on a Nextseq 500 high output flowcell and sequenced on a NextSeq 500 platform (Illumina) with 2×75bp paired-end chemistry.