Journal: Stem Cell Research & Therapy

Article Title: A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro

doi: 10.1186/s13287-023-03583-2

Figure Lengend Snippet: Expression of lineage markers in embryos cultured in vitro. A Immunofluorescence staining of Oct4 and Nanog from E5.5 embryos in the IVCM or IVCM + Cells group. Cells in the IVCM + Cells group are referred to as hPPSCs and hUVECs; scale bars, 200 µm. n = 11–23 for each group, P < 0.05 indicated statistical significance. B Immunofluorescence staining of Cdx2,Gata4 and Sox17 on E5.5 embryos in the IVCM or IVCM + Cells group; scale bars, 200 µm. C Immunofluorescence staining of Cdx2,Gata4,Sox17 and T on E6.5 embryos in the IVCM or IVCM + Cells group; scale bars, 200 µm

Article Snippet: The primary antibodies for embryo immunofluorescence were rabbit anti-OCT4 polyclonal antibody (1:200, Thermo Fisher Scientific, PA5-27438), rabbit anti-Nanog antibody (1:100, Thermo Fisher Scientific, PA5-85110), Rabbit anti-Cdx2 antibody (1:1000, Abclonal, A19030), Goat anti-Gata4 polyclonal antibody(1:100, R&D system, BAF2606), Goat anti-Sox17 polyclonal antibody(1:100, R&D system, AF1924), Goat anti-T/Brachyury polyclonal antibody(1:100, R&D system, AF2085).The secondary antibody was FITC-goat anti-rabbit IgG (1:500, Thermo Fisher Scientific, F-2765), Donkey anti-goat IgG H&L (Alexa Fluor® 488) (1:500, Abcam, ab150129).

Techniques: Expressing, Cell Culture, In Vitro, Immunofluorescence, Staining

Journal: International journal of molecular sciences

Article Title: Development of Human Adrenocortical Adenoma (HAA1) Cell Line from Zona Reticularis.

doi: 10.3390/ijms24010584

Figure Lengend Snippet: Figure 1. (A) HAA1 cells in culture. (B) Both master-regulator NR5A1/SF1 (green) and the key adrenal transcription GATA4 and GATA6 proteins (red) are expressed at a lower level in the HAA1 cell line (bottom panels) than in the control NCI-H295R (top panels) human adrenal cortical carcinoma cell line. Scale bar, 50 µ. (C) A representative Western blot analysis of protein expression; NR5A1/SF1 (top panel) and ACTIN1 (bottom panel); L1. SW13 adrenal carcinoma cells (these cells produce no steroids and serve as a negative control) L2. NCI-H295.L3. HAA1 (D) Quantitative analysis of GATA4 and GATA6 protein expression for the experiment shown in (B); *, p < 05; ***, p < 0.001. (E) Quantitative analysis of NR5A1/SF1 expression for the experiment shown in (C). All differences are significant, p < 0.05.

Article Snippet: The following antibody combinations were used: goat anti-GATA4 antibody (R&D Systems, Minneapolis, MN, USA), followed by donkey anti-goat Alexa Fluor 555-conjugated antibodies (Alexa); rabbit anti-GATA6 antibody (Cell Signaling Technology, Danvers, MA, USA) and rabbit anti-GLI1 antibody (Santa Cruz Biotechnology, Dallas, TX, USA) followed by goat anti-rabbit Alexa Fluor 488-conjugated antibodies (Invitrogen); and goat anti-SHH antibody, goat anti-HSD3B2, and goat anti-CYP11A1 (all Santa Cruz Biotechnology) antibodies followed by donkey anti-goat Alexa Fluor 555-conjugated antibodies (Invitrogen).

Techniques: Control, Western Blot, Expressing, Negative Control