Single-cell RNA sequencing of the liver cells from 2-month-old A/A and S/S mice with or without TNF challenge.

To gain insights into the liver transcriptomic profiles, we performed single-cell RNA sequencing of the liver single cells isolated from four groups of 2-month-old mice that included: A/A and S/S mice at baseline (n = 2 mice/genotype), and A/A and S/S mice after TNF challenge (n = 2 mice/genotype). TNF challenge (500 ng/mouse) is an established method to induce vaso-occlusive episode in S/S mice. The experiment goal is to dissect the effects of genotype and TNF challenge on the liver transcriptomic profiles. Therefore, four groups (total n = 8) of 2-month-old mice were included: A/A and S/S mice at baseline (n = 2 mice/genotype), and A/A and S/S mice after TNF challenge (n = 2 mice/genotype). For each group, the liver single cells of two mice were pooled together for single-cell RNA sequencing analysis. For TNF challenge, each mouse was intraperitoneally injected with 500 ng TNF and liver single cells were collected at 3 h after injection.