Fig. 4: PSMA is a SGSPC differentiation marker.

A Design of 2D CFC and 3D-matrigel organoid Matrigel experiments to assess clonogenicity of PSMA- and PSMA+ cells. B PSMA- cells demonstrate clonogenic capacity in 2D CFC and 3D-matrigel organoid assays, while PSMA+ cells do not. C PSMA- cells purified from 15 salivary gland patient samples, unlike PSMA+ purified cells, generate organoids in 3D culture. D Proposed hierarchical lineage model by which PSMA+ cells differentiate from the more primitive, stem/progenitor-like, self-renewing PSMA- cells. E Experimental design of 2D and 3D FACS-based characterization of growing PSMA- salivary cells to gauge PSMA+ cell generation. F Agarose gel (2%) image showing expression of FOLH1(PSMA, 182 bp) in two consecutive 3D organoid cultures but not in 2D SMG and PG cultures by RT-PCR. Uncropped gel images are supplied as Supplementary Fig. S12. G PSMA- cells plated in 2D culture generate PSMA+ cells at confluence. H 3D-matrigel organoids derived from PSMA- cells give rise to PSMA+ cells through multiple passages. I Illustration of CpG islands in human PSMA promoter region. J Schematic workflow of bisulfite conversion of 35 of 38 CpG, PCR and sanger sequencing of DNA obtained from SMG. K Hypermethylation ( ~ 70%) of CpG in PSMA promoter in SMG tissue. Each circle indicates individual CpG dinucleotides. White and dark circles represent unmethylated and methylated CpG, respectively.