Supplementary Figure 1.

Method for isolating HSCs from the livers of obese mice and mice with obesity-induced HCC. (A‒C), During the Nycodenz gradient separation, 6% to 17% Nycodenz gradient medium (0.06–0.17 g/mL) was used for further experiments and purification of HSCs. The purified cells in the white layer from NT and T segments were subjected to qPCR analysis for Lrat (A), Clec4f (B), or Stab2 (C) expression. Gene expression was normalised to GAPDH expression, and data are presented as fold-change difference (n = 6, 13 mice were used in 6 experiments). The average of mouse body weight was 57.6 g (range, 44‒67g). The raw data are also shown in the table. (D), Nycodenz gradient medium composition. E, Flow cytometry gating strategy. F, Isolated cell number from 10-, 35-, and 90-week-old mice. The body weight of mice: 10 weeks, 25.9 g; 35 weeks, 31.5 g; 90 weeks, 30.2g. The isolated cell number is shown in the table. Statistical significance was determined using Bonferroni's multiple comparisons test (see the raw data table by the bar graph).