Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: HNF4α Acts as Upstream Functional Regulator of Intestinal Wnt3 and Paneth Cell Fate

doi: 10.1016/j.jcmgh.2022.11.010

Figure Lengend Snippet: Human primary mesenchymal cells rescue Hnf4a Δ IEC jejunal growth in culture. ( A ) Crypts from control and Hnf4a Δ IEC mice were maintained in culture, with or without human mesenchymal cells. They were kept under culture for 5 days, and images were acquired each day using a Zeiss Celldiscoverer 7. Representative images of 2 independent technical replicates at 2 different original magnifications (2.5× and 20×) are shown. Scale bar = 500 μm for 2.5× magnification and 50 μm for 20× magnification. ( B ) Live co-culture sample stained at day 5 using CellMask Deep Red Plasma Membrane Stain ( red ) and Hoechst 33342 ( blue ). Three-dimensional imaging was performed using confocal microscopy. One enteroid surrounded by mesenchymal cells is shown. White dashed box emphasizes a mesenchymal cell that displays elongated telopods in close contact with epithelial cells ( white arrows ). ( C ) Areas (pixel ) of jejunal enteroid size were assessed using ImageJ and expressed as percentage of control enteroids (∗∗∗∗ P ≤ .0001). Mixed-species RNA-seq data obtained were analyzed using GSEA software for comparison with Paneth cell gene signature ( D ), goblet cell and tuft cell gene signatures ( E ), ISC gene signature ( F ), as well as differentiated and progenitor enterocyte gene signatures ( G ).

Article Snippet: First, 5-day-old co-cultures were labeled with Hoechst 33342 (Cell Signaling Technology, New England Biolabs) for 15 minutes at 37°C, followed by incubation with CellMask Deep Red Plasma Membrane Stain (Invitrogen) for 10 minutes at 37°C.

Techniques: Co-Culture Assay, Staining, Imaging, Confocal Microscopy, RNA Sequencing Assay, Software

Journal: PLoS ONE

Article Title: Zinc Oxide Nanoparticles Induce Necrosis and Apoptosis in Macrophages in a p47phox- and Nrf2-Independent Manner

doi: 10.1371/journal.pone.0065704

Figure Lengend Snippet: Representative images are shown for activated caspase-3 (first column) and corresponding DNA staining with Hoechst 33342 (second column). To control for unspecific staining, CC-3 antibody was substituted by IgG antibody (third column) and shown with the corresponding Hoechst 33342 staining (fourth column). Original magnification 400×.

Article Snippet: Nuclear staining was performed by using the dye Hoechst 33342 (Cell Signaling) in a final concentration of 1 µg/ml in PBS for 15 min at 37°C in a humid box.

Techniques: Staining