Fig. 4: Navitoclax and Venetoclax in the cerulein-induced model of CP.

A Schematic representation of the human pancreatic stellate cell (hPSC) activation model. Created with BioRender. B Relative ACTA2 mRNA expression in hPSCs at different activation stages and following treatment with 100 nM Navitoclax or Venetoclax. Data are presented as fold change relative to quiescent hPSCs (qhPSCs) (N = 3). Values represent mean ± SD. Statistical significance was determined using one-way ANOVA with Sidak’s post hoc test. C Cell death in hPSCs with different activation states after 48 hour treatment with 100 nM Navitoclax or Venetoclax, shown as the percentage of dead cells relative to total cell number. Data from 4 independent experiments (N = 4); 8 images per condition. Total cell counts: qhPSC: 3335, qhPSC + Ven: 3705, qhPSC + Navi: 3292, shPSC: 1140, shPSC + Ven: 1117, shPSC + Navi: 1146, ahPSC: 1241, ahPSC + Ven: 1107, ahPSC + Navi: 1141. Results are shown as mean ± SD. Statistical analysis performed using one-way ANOVA with Sidak’s post hoc test. D Representative images of hematoxylin and eosin (H&E) stained mouse pancreata from the cerulein-induced CP model. Tissue injury and atrophy are particularly evident in the groups with induced CP, especially with additional administration of Navitoclax. Scale bar: 100 µm. E Heatmap depicting the individual parameters included in the histological evaluation of tissues from the CP model, based on H&E staining. Legend: Parenchymal integrity (Par, 0/3/5/7), inflammatory infiltration (Infltr, 0/1/2/3), acinar atrophy (AcAt, 0/3/5/7), pseudotubular complexes (PsTub, 0/1/2/3), and fibrosis (Fibr, 0/3/5/7). Adapted from previous publications and modified [20, 25]. F Overall histological severity score (0−27) for each group from the CP model, based on H&E staining. Results presented as mean ± SD. Statistical analysis was performed using the non-parametric Kruskal-Wallis test followed by Dunn’s post hoc test. G Representative images of Sirius Red staining of mouse pancreata from the cerulein-induced CP model, which highlights total collagen in the tissue. Increased fibrosis is notable across all groups with induced CP, particularly significant upon additional administration of Navitoclax. Scale bar: 100 µm. H Percentage of tissue area from the CP model represented by total collagen, calculated across entire tissue scans from Sirius Red-stained sections. Results are presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Sidak’s post hoc test. I Mass of pancreata from the CP model, weighed at postmortem dissection. Results are presented as mean ± SD. Statistical analysis was performed using the non-parametric Kruskal-Wallis test followed by Dunn’s post hoc test. J, K Enzymatic activity measurements of lipase (G) and amylase (H) from mouse plasma in the CP model. Measurements at 18 h post the first cerulein administration (18 h), after the 4th week (4 wk), and postmortem at the end of the 8th week (8 wk). Results are presented as mean ± SD. Statistical analyses for lipase at 8 weeks were performed using one-way ANOVA followed by Sidak’s post hoc test, and for amylase at 8 weeks, the Kruskal-Wallis test followed by Dunn’s post hoc test For earlier time points, unpaired t-tests with Welch’s correction were applied. For the cerulein-induced CP model, n = 6 for all groups, except for the CP Navi group with n = 7. Figure Abbreviation Legend: Sham (saline + vehicle), Navi (saline + Navitoclax), Ven (saline + Venetoclax), CP (cerulein + vehicle), CP + Navi (cerulein + Navitoclax), CP + Ven (cerulein + Venetoclax).