The reliability of colorimetry is precise(ly) as expected.

Figure S6. Gene ontology (GO) analysis of the differentially expressed genes between C1 and C2. (JPG 403 kb

Additional file 1: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Table S1. Primary and secondary antibodies used for immunofluorescence or FACS analysis. (DOCX 15 kb

Additional file 3: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S1. Enrichment of porcine male germline stem cell (mGSCs). (a) The testicular suspension contained several cell types. (b) Testicular fibroblast cells (TFCs) were removed by differential attachment technique. (c) The unattached cells underwent density gradient centrifuging to remove the leukomonocytes (LYMs) and cell debris (d) and red blood cells (RBCs) (e), and to retain mGSCs (f). (JPG 699 kb

Additional file 5: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S3. Cell cycle assay of putative porcine male germline stem cell (mGSCs). Cell cycle analyses at cell culture days 3, 5, and 7 revealed that only a small percentage of the putative mGSCs entered the S and G2 cell division phases. Meanwhile, part of putative mGSCs underwent apoptosis detected by analysis of apoptosis at day 7 of cell culture. (JPG 285 kb

Additional file 10: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S8. Induction of C2 clusters differentiation into mature adult Leydig cells (ALCs). The C2 clusters began to differentiate into fibroblast cells at day 1 (a), became smaller from days 3 to 6 (b, c), and had disappeared by day 9 (d) The C2 clusters were fully differentiated into ALCs by day 12 (e), with abundant mature ALCs observed by day 15 (f). (JPG 558 kb

Additional file 4: of Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Figure S2. Immunolocalization of VASA and NANOG proteins in the testis of 5-day-old piglet. Immunohistochemistry analysis showed that the reproduction-associated marker VASA localized to the basement membrane of seminiferous tubules, whereas the pluripotent factor NANOG was expressed in the interstitial space and seminiferous tubules. (JPG 442 kb