Journal: Stem Cells Translational Medicine
Article Title: EPHA2 is a novel cell surface marker of OCT4-positive undifferentiated cells during the differentiation of mouse and human pluripotent stem cells
doi: 10.1093/stcltm/szae036
Figure Lengend Snippet: Co-expression of EPHA2 with OCT4 in EBs during human iPSC differentiation into hepatocyte. (A, B) Relative gene expression of undifferentiation and differentiation markers during hepatic induction. Statistical analysis was done by Dunnett’s test against day 0 and graphed as means ± SE of 3 independent biological replicates. * P < .05; ** P < .01, *** P < .001. (C) Immunofluorescent staining of EBs at days 5, 8, and 10. Bars; 200 μm. (D) Enlarged images of EB at day 5 in (C). Bars; 50 μm. (E, F) Quantification of immune-positive cells in . Box plot showing the percentage of EPHA2 + cells among SOX17 + or OCT4 + cells (E). Box plot showing the percentage of EPHA2 + and TRA1-81 + cells among OCT4 + cells (F). Each box represents 1st quartile, median, and 3rd quartile, and whiskers show the minimum and maximum values. Ten images of independent EBs were analyzed. Total count of DAPI + nuclei per image were between 1 × 10 3 and 2 × 10 3 . Statistical significance was defined by Tukey test of SOX17 and OCT4, respectively in (E) and t -test between EPHA2 and TRA1-81 in (F). **P p < .01, *** P < .001, N.S; no significance between 3 with P > .05.
Article Snippet: For immunofluorescent staining, the following primary antibodies were used: rabbit polyclonal anti-mouse and human OCT4 (sc-9081; Santa Cruz Biotechnology), mouse monoclonal anti-OCT4 (sc-5279; Santa Cruz Biotechnology), rabbit polyclonal anti-mouse NANOG (RCAB0001P; REPROCELL), mouse monoclonal anti-mouse SSEA-1 (TM13; Kyowa Medex), rabbit polyclonal anti-mouse EPHA2 (sc-924; Santa Cruz Biotechnology), mouse monoclonal anti-β-TUBULIN (T5293, Sigma), goat polyclonal anti-mouse AFP (sc-8108; Santa Cruz Biotechnology), and goat anti-mouse and human ALB (55727; MP Biomedicals).
Techniques: Expressing, Gene Expression, Staining