sc 9081  (Santa Cruz Biotechnology)

Journal: Stem Cell Research & Therapy

Article Title: SMYD1 modulates the proliferation of multipotent cardiac progenitor cells derived from human pluripotent stem cells during myocardial differentiation through GSK3β/β-catenin&ERK signaling

doi: 10.1186/s13287-024-03899-7

Figure Lengend Snippet: Primary and secondary antibodies

Article Snippet: Primary , Anti-OCT4 , Santa Cruz sc-9081 , Mouse , Immunofluorescence , 1:100.

Techniques: Immunofluorescence, Western Blot, Flow Cytometry, ChIP-chip

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: Expression of EPHA2 in heterogenous subpopulation of human PSCs. (A) qRT-PCR analysis of EPHA2 and undifferentiated state-specific marker genes during EB-based random differentiation of human iPSCs (201B7) without basic FGF. Statistical analysis was done by Dunnett’s test comparing to day 0 and graphed as means ± SE of 3 independent experiments. * P < .05; ** P < .01. (B) Representative flow cytometric plots of living human iPSCs stained with EPHA2 antibody-conjugated with AFF488. Gate3 and Gate4 were sorted as EPHA2 − and EPHA2 + cell populations, respectively. See also . (C) Feature plots of EPHA2 and undifferentiated state-specific genes in publicly available undifferentiated human ESC H1 and H9 data subsets from GSE75748 scRNA-seq dataset. Note that EPHA2 expression in hESCs was heterogeneous. Normalized expression levels were plotted. (D, E) Immunofluorescent staining of human iPSC cultured on SyntheMax II-coated plate with StemFit medium. The cells were fixed with paraformaldehyde in PBS and permeabilized. Bars; 200 μm. (F) qRT-PCR analysis of fractioned EPHA2 + and EPHA2 - subpopulations. EPHA2 + cells express higher OCT4 and NANOG , than EPHA2 − cells. Means ± SE of 3 independent experiments were shown. Statistical significance was defined as * P < .05 by t -test.

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, Quantitative RT-PCR, Marker, Staining, Cell Culture

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: Transplantation of EPHA2 + cells into immune-deficient mice formed tumors in vivo. (A) Immunofluorescent staining of mouse EBs differentiated into hepatocyte lineages. Expression of an early hepatocyte marker AFP at day 10 and a mature marker ALB at day 14 were detected. Bars; 200 μm. (B) Depletion of undifferentiated ES colonies after removal of EPHA2 + cells from EBs. Oct4-egfp ESCs were differentiated by EB formation for 10 and 14 days. EPHA2 + cells were removed from EBs using anti-EPHA2 antibody-bound MACS after dissociation with trypsin/EDTA. The residual cells were cultured in ES maintenance medium with LIF for 7 days. The alkaline phosphatase (AP) activity was visualized by incubating with AP substrate. Bar; 2 cm. (C) The number of EGFP + cell colonies in (B). Statistical analysis was done by Tukey test and graphed as means ± SE of 4 independent experiments. (D) Scheme of in vivo transplantation experiment. Mouse ESCs (D3) were differentiated into hepatocyte linages and the EBs were dissociated by EDTA treatment. The cells were transplanted into SCID mice after depletion of EPHA2 + cells by MACS. (E) Decreased teratoma formation after transplantation of EPHA2 − cells. White arrowheads indicate teratomas. (F) H&E staining of teratomas formed in the testicular subcutaneous tissue without MACS procedure. Typical cell types of 3 germ layers were confirmed. Bar; 500 μm. (G) Typical teratoma formation after transplantation of dissociated EB at day 10 through hepatic portal vein. White arrowheads indicate teratomas. (H) Quantification of teratoma formation in (E) and (G). Statistical analysis was done by Chi-square test, * P < .05, ** P < .01.

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: Transplantation Assay, In Vivo, Staining, Expressing, Marker, Cell Culture, Activity Assay

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