Fig. S3.

Role of rhHMGB1 in bulk pancreatic cancer cell stemness and proliferation. (a) Flow cytometric analysis of the percentage of CD133⁺ cancer cells among bulk cells at 72 h post-treatment with 150 ng/mL rhHMGB1. (b) CCK8 testing was performed to assess the ability of CD133⁺ cancer cells to proliferate when co-cultured with the following agents: i) PBS; ii) irradiated HMGB1⁺ cells; iii) irradiated HMGB1⁻ cells (HMGB1-shRNA1); iv) rhHMGB1 (150 ng/mL) for 0, 2, 4, or 6 d. (c) Flow cytometric analysis of cell cycle progression in CD133⁺ cancer cells at 48 h post-treatment with 150 ng/mL rhHMGB1. (d) Colony-forming assay analysis of the colony forming ability at 48 h post-treatment with 150 ng/mL rhHMGB1. Experiments were repeated three times, and the data are expressed as the mean ± SEM.