Fig 2: Expression of Gli2 and PD-L1 in huTGO lines and patient tumor tissue(A) Expression of Gli2 and PD-L1 in TGO lines generated. Immunofluorescence images for the expression of PD-L1 (green) and Gli2 (red) in (B, C) huTGO2, and (D, E) huTGO7. (F) Quantification of proliferating cells within huTGO2 and huTGO7 organoid lines. *P < 0.05 compared to huTGO2, n = 4 individual organoids. Immunofluorescence staining for Gli2 (red) in patient tumor tissue from which (G) huTGO2 and (H) huTGO7 were derived. Immunohistochemistry for the expression of PD-L1 in patient tumor tissue from which (I) huTGO2 and (J) huTGO7 were derived. (K) Proliferation in patient tumor tissue from which huTGO2 and huTGO7 were derived. *P < 0.05 compared to tumor tissue from patient from which huTGO2 was derived, n = 6 individual high power fields.

Fig 3: Gli2 and PD-L1 expression in gastric cancer organoids derived from iLgr5;GLI2A miceImmunofluorescence staining for proliferating EdU (green), Gli2 (red) and PD-L1 (cyan) expression in organoids derived from control normal (mFGO, A–D) and tumor iLgr5;GLI2A-derived (mTGO, E–H) mouse tissues. (I) Immunofluorescence images of PD-L1 (red) and EdU (green) expression in mTGOs, treated with 0, 1, 5, or 10 µM GANT61. (J) Quantification of % EdU positive cells/total cells in GANT61 treated mTGOs. *P < 0.05 compared to 0 µM GANT61, n = 4 per group. (K) Western blot analysis for the expression of PD-L1 using whole cell lysates collected from mTGOs, treated with 0, 1, 5, or 10 µM GANT61. (L) Quantification of western blots expressed as the PD-L1/GAPDH average fold change relative to 0 µM GANT61. *P < 0.05 compared to 0 µM GANT61, n = 4 per group.