Labchip gxtouchht system

Monocytes were cultured for 3 d in the presence of 100 ng ml⁻¹ of M-CSF, 5 ng ml⁻¹ of IL-4 and 20 ng ml⁻¹ of TNF. Total RNA was extracted using the RNAeasy minikit (QIAGEN) including on-column DNase digestion according to the manufacturer’s protocol. The integrity of the RNA was confirmed in BioAnalyzer using RNA 6000 Pico kit (Agilent Technologies) (8.8 < RNA integrity no. (RIN) < 10). Libraries were prepared according to Illumina’s instructions accompanying the TruSeq Stranded mRNA Library Prep Kit (Illumina). RNA, 500 ng, was used for each sample. Library length profiles were controlled using the LabChip GXTouchHT system (Perkin Elmer). Sequencing was performed in four sequencing units of NovaSeq 6000 (Illumina) (100–593 nt length reads, paired end) with an average depth of 40 × 10⁶ clusters per sample. RNA-seq data discussed in this publication have been deposited in the NCBI’s GEO and can be accessed at accession no. GSE188982.

Epidermal cells were isolated 6 hr after vehicle or papain treatment. Cells were lysed in RLT buffer (QIAGEN). Total RNA was extracted using the RNAeasy minikit (QIAGEN) including on-column DNase digestion according to the manufacturer’s protocol. The integrity of the RNA was confirmed in BioAnalyzer using RNA 6000 Nano kit (Agilent Technologies) (8.8 < RIN < 10). Libraries were prepared according to Illumina’s instructions accompanying the TruSeq Stranded mRNA Library Prep Kit (Illumina). 500 ng of RNA was used for each sample. Library length profiles were controlled with the LabChip GXTouchHT system (Perkin Elmer). Sequencing was performed in three sequencing unit of NovaSeq 6000 (Illumina) (100-nt-length reads, paired end) with an average depth of 40 millions of clusters per sample.