Skip to main content
. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Cancer Discov. 2021 Jan 15;11(6):1562–1581. doi: 10.1158/2159-8290.CD-20-1109

Figure 3. ARID1A knockout induces mucinous metaplasia.

Figure 3.

A, Schematic illustration of gastric epithelium. Different cell lineages and specific lineage markers are indicated. B, Western blot of mucin-producing pit cell and mucous neck cell markers, TFF1, TFF2 and LYZ, reveals upregulation in TP53/ARID1A DKO organoids. Quantification of expression from independent experiments (N>3) was shown. Dots indicate independent experiments. C, Immunofluorescence staining of MUC5AC (green) and TFF1 (red) in engineered organoids and the donor primary gastric tissues. Nuclei were stained with DAPI (blue). Quantification of MUC5AC-positive organoids is shown. D, Mucin production in engineered organoids and donor primary gastric tissues detected by Alcian blue staining. Nuclei were counterstained by nuclear fast red. Quantification of Alcian blue-positive organoids indicate increased mucin in all five TP53/ARID1A DKO organoids lines. E, TP53/ARID1A DKO xenografts in a subcutaneously xenografted NSG mice retain their mucin-secreting phenotype in vivo. Alcian blue and PAS staining. Goblet-like (Alcian blue -positive) and pit-like (PAS positive) cells were indicated. F, Quantification of mitotic cells. Goblet-like and pit-like cells with mitotic figures were shown. H&E staining. G, Immunofluorescence staining of TP53/ARID1A DKO organoids showing LYZ-positive (red) or MUC5-positive (red) proliferating cells (KI67+, green). H, IHC staining of CDX2 in xenografts and the donor primary gastric tissues. Colon tissues were used as positive control. I, IHC staining of MUC2 in organoids, xenografts and the donor primary gastric tissues. Colon tissues were used as positive control. J, Immunofluorescence staining of CLDN18 (white), MUC6 (red) and PGC (green) in TP53/ARID1A DKO organoids, xenografts and the donor primary gastric tissues. Cells within SPEM features (MUC6 and PGC double positive) are marked by arrows. K, A significant negative correlation between ARID1A (brown) IHC expression and mucin (blue, Alcian blue) production was identified in a human gastric cancer tissue microarray (total 197 patients).