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[Preprint]. 2023 Feb 24:2023.02.23.529824. [Version 1] doi: 10.1101/2023.02.23.529824

Fig. 4. Tex10 overexpression enhances PGCLC specification efficiency.

Fig. 4.

(A) Cellular morphology of Tex10 overexpression (OE) vs. control EBs (Empty vector, EV) at D2, D4, and D6 PGCLC stages (scale bar, 100 μm).

(B) Flow cytometry analysis of the efficiency of PGCLC specification using the markers SSEA1 and CD61. Percentages of double-positive (SSEA1+ and CD61+) cells are indicated at day 6 of PGCLC induction. Two cell clones, OE1 and OE2, were used as biological replicates and the paired t-test was used to detect significance.

(C) Quantitative RT-PCR analysis of Tex10, Otx2, and Nanog expression during the transition from EpiLCs to D2, D4, and D6 EBs. ANOVA was performed for the significance test.

(D) Western blot analysis of FLAG-Tex10, Otx2, and Nanog expression during the transition from EpiLCs to D2, D4, and D6 EBs. Quantification values for Otx2 and Nanog relative to Actin are shown on the top bands for D6EB samples.

(E-G) Flow cytometry analysis of PGCLC specification efficiency using cell surface markers SSEA1 and CD61. Percentages of double-positive (SSEA1+ and CD61+) cells are indicated at day 6 of PGCLC induction with (+) or without (−) cytokines (E). Quantification of double-positive percentages in live cells (F) and numbers per 20,000 analyzed cells (G) are shown with bar plots. Two cell clones, OE1 and OE2, were used as biological replicates. The ANOVA test was used to detect significance.