Journal: Nature communications

Article Title: FOXO1 Inhibition Yields Functional Insulin-Producing Cells In Human Gut Organoid Cultures

doi: 10.1038/ncomms5242

Figure Lengend Snippet: Insulin-positive cells in 184-day-old human gut organoids. a–b , qPCR analysis of different markers in gut organoids transduced with control (empty bars) or HA-Δ256 FOXO1 adenovirus (black bars). c , Quantification of insulin- and GLP1-positive cells in gut organoids transduced with control (empty bars) or HA-Δ256 FOXO1 adenovirus (black bars). d–e , Immunohistochemistry with insulin (green), C-peptide (red), and CGA (magenta). f , Magnification of a typical flask-shaped insulin-positive cell from panel e . g–h , Co-immunohistochemistry with insulin (green), HA (to detect HA-Δ256 Foxo1 adenovirus) (red), and CGA (magenta). i–j , Co-immunohistochemistry with insulin (green) and FOXO1 (red), or ( k, l ) insulin (green), α-SMA (red) and CDX2 (magenta). Insets in h, j, and l show magnifications of individual cells. DAPI (blue) was used throughout to visualize DNA. Scale bars: 50 µm in a–e ; 10 µm in f (n=3–6 for qPCR and 3 for histology) (* p < 0.05). We present quantitative data as means ± SEM.

Article Snippet: We counterstained DNA with DAPI (Cell Signaling).

Techniques: Transduction, Control, Immunohistochemistry