Journal: iScience

Article Title: Nicotinamide mononucleotide restores impaired metabolism, endothelial cell proliferation and angiogenesis in old sedentary male mice

doi: 10.1016/j.isci.2024.111656

Figure Lengend Snippet:

Article Snippet: Rabbit anti-ERG , Cell Signaling Technology , Cat#Ab92513.

Techniques: Recombinant, Isolation, SYBR Green Assay, Membrane, Saline, Western Blot, Staining, Cell Culture, Imaging, Software, Functional Assay

Journal: International Journal of Molecular Sciences

Article Title: Bhlhe40 Regulates Proliferation and Angiogenesis in Mouse Embryoid Bodies under Hypoxia

doi: 10.3390/ijms25147669

Figure Lengend Snippet: The Bhlhe40 knockout prevents hypoxia-induced cell cycle arrest in mature endothelial cells (ECs) in vascular structures in mouse EBs. EBs from Bhlhe40-Ctrl mESC (−DOX) and Bhlhe40-KO mESC (+DOX) cell lines, generated from iBhlhe40 ΔSg4 R3.8 Cas9 mESCs, were differentiated during 10 days in normoxia (21% O 2 , N) or treated during the last 48 h in hypoxia (1% O 2 , Hx) and were pulse-labeled with EdU during the last 3 h: ( A ) A representative image of vascular structures of each experimental condition (N Bhlhe40-Ctrl (−DOX), N Bhlhe40-KO (+DOX), Hx Bhlhe40-Ctrl (−DOX), and Hx Bhlhe40-KO (+DOX)) is shown. Vascular structures were visualized by double immunostaining using anti-CD31 (red) and anti-ERG (green). S-phase cells were visualized by EdU incorporation (magenta). Bar: 50 µm. Magnifications of the areas marked with the white squares are displayed (third column). Bar: 40 µm. CD31 masks (grey line) are shown in the last column. ERG + EdU − nuclei (green triangles) and ERG + EdU + nuclei (white triangles) are indicated. ( B ) Percentage of EdU + ECs in vascular structures in EBs differentiated 10 days in normoxia (N, blue circles) or 48 h hypoxia (Hx, red triangles). 4–10 fields per EB were quantified out of a total of 6 EBs per experimental condition (1.5 × 10 3 –3 × 10 3 ECs quantified per experimental condition). Each symbol corresponds to one field, and the horizontal line represents the mean of all fields quantified per experimental condition. Statistical significance was determined by the Mann-Whitney test (ns = not significant; ** p < 0.01; *** p < 0.001).

Article Snippet: Antibodies for immunostaining were diluted in PBS with 3% BSA and 0.1% Tween-20 as follows: 1:800 rat anti-mouse CD31 (BD, Franklin Lakes, NJ, USA, 553370), 1:1000 rabbit anti-ERG (Abcam, Cambridge, UK, ab92513), 1:250 rabbit anti-GFP (Thermo Fisher Scientific, Waltham, MA, USA, A-6455) and 1:100 rabbit anti-mouse HOXD9 (Santa Cruz Biotechnology, Dallas, TX, USA, sc-8320); and 1:500 goat anti-rat AF564, 1:500 goat anti-rabbit AF488, and 1:300 goat anti-rabbit AF594.

Techniques: Knock-Out, Generated, Labeling, Double Immunostaining, MANN-WHITNEY