Journal: bioRxiv

Article Title: Genetic dysregulation of an endothelial Ras signaling network in vein of Galen malformations

doi: 10.1101/2023.03.18.532837

Figure Lengend Snippet: (a) Vascular defects in homozygous and heterozygous EPHB4 mutant embryos. Shown are images of littermate embryos of the indicated genotypes at E10.5. Note abnormal vascularization of the yolk sac, reduced size and distended pericardial sac of homozygous EPHB4 F867L mouse embryos. (b) Honeycomb-like vascular patterning in homozygous F867L EPHB4 mutants. E10.5 yolk sacs were stained with anti-CD31 antibodies to identify the vasculature. For the EPHB4 F867L yolk sac, the image right is a higher magnification of the boxed area in the left image. Note hierarchical vascular networks in WT and EPHB4 F867L heterozygous yolk sacs and primitive vascular plexuses in EPHB4 F867L homozygous yolk sacs. (c) Characterization of vascular defects E9.5 Ephb4 F 867 L/F867L embryos. CD31 antibody staining of littermate E9.5 embryos of the indicated genotypes. For each embryo, the right image is a higher magnification of the boxed area in the left image. Arrows show trunk angiogenesis toward the midline in EPHB4 F867L heterozygous but not EPHB4 homozygous embryos. (d and e) Embryos of the indicated genotypes were administered tamoxifen at E13.5 and harvested at the indicated time points. (d) MAPK activation assessment of E16.5 embryos by Western blotting. Activation of MAPK in liver lysates was determined by Western blotting using anti-pERK antibodies. Equal protein loading was determined by antibody probing for ERK and actin abundance. (e) Extensive cutaneous vascular hemorrhage in Ephb4 fl/F867L Cdh5ert2cre embryos. Gross images of E18 embryos (left) and images of tissue sections of skin from the same embryos stained with anti-CD31 antibody (right). Note hemorrhagic appearance and extravasation of green auto-fluorescent erythrocytes in Ephb4 fl/F867L Cdh5ert2cre embryos (arrows).

Article Snippet: Extracts were subsequently centrifuged at 13,000 x g for 10 minutes at 4 °C and pERK levels were determined by Western blotting as above, using rabbit anti-phospho-ERK monoclonal IgG (clone D13.14.E, Cell Signaling Technology, 4370) and rabbit anti-ERK IgG (clone 137F5, Cell Signaling Technology, 4695) and rabbit anti-ACTB polyclonal IgG.

Techniques: Mutagenesis, Staining, Activation Assay, Western Blot

Journal: Frontiers in Microbiology

Article Title: The first Brevinin-1 antimicrobial peptide with LPS-neutralizing and anti-inflammatory activities in vitro and in vivo

doi: 10.3389/fmicb.2023.1102576

Figure Lengend Snippet: Effects of Brevinin-1GHd on inflammatory pathway activation inspired by LPS. (A) Typical WB bands of ERK, p38, JNK in RAW 264.7 cells. The cells were pre-treated with or without 1, 2, and 4 μM Brevinin-1GHd for 30 min and an additional 30 min after LPS addition, then were collected for WB analysis. ICONS 1–5 are the control group, LPS treatment group, and 100 ng/ml LPS plus 1, 2, and 4 μM Brevinin-1GHd treatment groups, respectively. (B) Quantification analysis of band densities in A figure. Data are expressed as mean ± SEM ( n = 3). ∗∗ p < 0.01 and ∗∗∗ p < 0.001 as well as ### p < 0.001 suggest significant difference in comparison with the control incubated with 100 ng/ml LPS and with the control without both LPS and Brevinin-1GHd, respectively.

Article Snippet: Then, the macrophages were treated with different concentrations of Brevinin-1GHd (1, 2, and 4 μM) for 30 min, and then co-incubated with 100 ng/ml LPS for another 30 min before collected for WB analysis of phospho-JNK/JNK, phospho-ERK/ERK, phospho-p38/p38 and GAPDH contents (Cell Signaling Technology, Beverly, Massachusetts, USA) as previously described ( ).

Techniques: Activation Assay, Incubation