Mitochondrial perturbation in the intestine causes microbiota-dependent injury and gene signatures discriminative of inflammatory disease
Mitochondrial perturbation in the intestine causes microbiota-dependent injury and gene signatures discriminative of inflammatory disease
The entire colon was removed from either Hsp60Δ/ΔIEC or Hsp60fl/fl mice weighed and stored in Dulbecco's Modified Eagle Medium (DMEM). After being washed with PBS and minced on ice, tissue specimens were dissociated and enzymatically digested (Multi Tissue Dissociation Kit using gentleMACS™ Dissociator, Miltenyi Biotec). After centrifugation and resuspension in PBS+2% FCS, the cell suspension was strained through a 70-µm strainer, spun down and resuspended in 2% FCS/PBS including RNase inhibitor (New England Biolabs, catalog no. M0314L,1:100). Debris removal solution (Miltenyi Biotec, catalog no. 130-109-398) was used to remove cell debris from the dissociated tissue. Then the dead cell removal kit (Miltenyi, catalog no. 130-090-101) was used to enrich live cells. CD45+ cells were sorted out using the CD45 Microbeads kit (Miltenyi Biotec, catalog no. 130-052-301).  https://cdn.10xgenomics.com/image/upload/v1660261285/support-documents/CG000204_ChromiumNextGEMSingleCell3_v3.1_Rev_D.pdf https://cdn.10xgenomics.com/image/upload/v1660261285/support-documents/CG000204_ChromiumNextGEMSingleCell3_v3.1_Rev_D.pdf