Figure 4: Stem cell- and germ cell-specific marker expression and the doubling time analysis in PC and GFP PGC lines. 4 female PC PGC lines (FS111, FS214, FS302, and FS319), 4 male PC PGC lines (FS101, FS117, FS202, and FS210), 1 female GFP PGC line (5ZP), and 3 male GFP PGC lines (4ZP, 67ZP, and 8ZP) were investigated in detail. We examined two biological samples as repeats. 3 parallel samples were used at qPCR runs. We analysed the CVH (a), cDAZL (b), cPOUV (c), and cNANOG (d) (using cGAPDH as reference gene) and the expression of a stem cell-specific miR-302a (relative to miR-92 as reference gene). Gene expression values were calculated using GenEx software relative to the FS101 sample in each case. Supplementary Table 1 contains detailed information of the analysed genes and the sequences of the primers used at qPCR runs. (f) Furthermore, the proliferation rate of the cell lines was measured on 3 different days using CCK-8 proliferation assay. We used three 96-well plates with 6-6 parallel wells. The doubling time was calculated according to the measured optical densities (OD). PC PGC lines: Partridge colour Hungarian chicken embryo-derived primordial germ cell lines. GFP PGC lines: green fluorescent protein- (GFP-) expressing transgenic chicken embryo-derived primordial germ cell lines.