Sample 2
Mus musculus
Protocols: Colon fibroblasts were isolated from tdTomatoColVI reporter mice. Colon tissue was digested with DNase I, Collagenase D and Dispase I for 15minutes at 37°C. Murine fibroblasts were cultured with DMEM F12 (10% FCS, 1% Pen/Strep) and separated by fluorescence-activated cell sorting (FACS) on the basis of the expression of tdTOMATO. tdTOMATO+ and tdTOMATO- fibroblasts were stimulated with IL-6 or IL-11 (100 ng/mL) or PBS for 4 h. RNA isolated was performed according to the manufacturer's protocol (Macherey-Nagel, Nucleo Spin RNA) including an additional DNAse digestion step as recommended. Libraries were constructed Illumina Stranded mRNA Kit