Cold ischemia 48 hours
One kidney section from each timepoint was placed on 10x Visium FFPE (formalin fixed paraffin embedded) slide using CytAssist instrument. Adjacent sections (20 mm) were used for RNA quality assessment and samples with RQN value 7 were used for spatial transcriptomic analysis. Tissue processing and downstream analyses were achieved by adopting the following 10x Genomics protocols without any alteration: (1) Visium CytAssist Spatial Gene Expression for FFPE (CG000520, Revision B) for deparaffinization, staining, decrosslinking and imaging of slides. (2) Sequencing sections of the Visium CytAssist Spatial Gene Expression Reagent Kits User Guide (CG000495) for hybridization. The CytAssist libraries were sequenced using a Novaseq 6000 sequencer. Gene expression counts and spot positions were obtained using the SpaceRanger pipeline (v2.0.1) for downstream analysis.