Fig 2: Aged gastric organoids express Tbx3. A, The expression of Wnt target genes in young and aged gastric organoids (n = 3). B, Immunofluorescence for Tbx3 in young and aged gastric organoids (n = 5). E-cad: E-cadherin. C,Mki67 and Tbx3 expression in the presence (WRC+) and the absence (WRC-) of Wnt3a, R-Spondin1, and Chir99021 in aged organoids (n = 3). D, Western blot analysis for TBX3 and ß-actin of AGS cells transfected with empty vector or TBX3-expressing plasmid. E, MTS assay for TBX3-overexpressing AGS cells (n = 3). F, Representative images of SABG assay for TBX3-overexpressing AGS cells. Scale bar: 50 µm. G, The ratio of senescent cells in TBX3-overexpressing AGS cells assessed by SABG assay (n = 3). n.s., not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Fig 3: SP and ECAD expression and localization was severely affected in MSG sections from pSS and sicca patients. Representative images of double immunofluorescence staining of substance P (SP, red) and e-cadherin (ECAD, green) in pSS and sicca patients MSG sections. Scale bar 25 µm; 40× magnification.

Fig 4: Cellular senescence is suppressed in aged gastric organoids. A, GSEA for (left) G2–M checkpoint-related genes and (right) senescence-related genes. B, The expression of genes related to cellular proliferation and cell cycle in young and aged gastric organoids (n = 3). C, The expression of senescence-inducing genes in young and aged gastric organoids (n = 3). D, Immunofluorescence for genes related to cellular proliferation and senescence in young and aged gastric organoids (n = 5). E-cad: E-cadherin. E, Representative images of SABG assay for young and aged gastric organoids. Scale bar: 50 µm. F, The ratio of senescent cells in young and aged gastric organoids assessed by SABG assay (n = 3). G, The expression of SASP-related genes in young and aged gastric organoids (n = 3). n.s., not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.