Colonic epithelial cells isolated from WT and NOX1KO littermates by BD Melody sorter from proximal colon or distal colon regions were pooled separately across 2 mice at equal ratios and were resuspended in 1% BSA in PBS and counted with trypan blue staining for cell viability and number. Approximately 30,000 cells per sample were loaded onto separate lanes of the Chromium X Controller. Gene expression libraries were prepared according to manufacturer instructions using the 10X Genomics Single Cell 3' Reagent Kit (v3.1 Chemistry Dual Index).