Journal: Frontiers in Cardiovascular Medicine

Article Title: Deficiency of Myeloid Pfkfb3 Protects Mice From Lung Edema and Cardiac Dysfunction in LPS-Induced Endotoxemia

doi: 10.3389/fcvm.2021.745810

Figure Lengend Snippet: Myeloid-specific Pfkfb3 deficiency attenuates LPS-induced inflammatory responses. (A) Representative images (left) and quantification (right) for immunohistochemical staining of the neutrophil marker Ly6G in lung sections of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 5). (B) Representative images (left) and quantification (right) for immunohistochemical staining of the macrophage marker Mac2 in lung sections of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 5). (C–E) qPCR analysis of the mRNA levels of Il1b (C) , Il6 (D) and Nos2 (E) in the lung of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 6). (F,G) ELISA analysis of Il1b (F) and Il6 (G) in serum of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 4). (H) NO levels in serum of Pfkfb3 WT mice and Pfkfb3 ΔMϕ mice 6 h after LPS injection ( n = 3–4). All data are represented as mean ± SEM, ** P < 0.01 and *** P < 0.001 for Pfkfb3 WT vs. Pfkfb3 ΔMϕ (unpaired two-tailed Student's t test).

Article Snippet: After antigen retrieval with Antigen Unmasking Solution (H-3301, Vector Laboratories, Burlingame, CA, USA) at 98°C for 10 min, sections were blocked with avidin solution with 10% normal rabbit serum for 1 h at room temperature, and incubated in biotin blocking solution with primary antibodies against Mac2 (3 μg/mL, CL8942F, Cedarlane, Burlington, NC, USA), or Ly6G (3 μg/mL, 551459, BD biosciences, San Jose, CA, USA) at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Marker, Injection, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: The Journal of Clinical Investigation

Article Title: ADAMTS7 promotes smooth muscle foam cell expansion in atherosclerosis

doi: 10.1172/JCI187451

Figure Lengend Snippet: ( A ) Schematic outlining experimental design and the generation of the SMC transgenic ADAMTS7 mouse. Created with BioRender.com. ( B ) ORO staining of the en face aorta and quantification of lesion area. n = 9. ( C ) Representative images of H&E-stained aortic root sections, quantification of plaque areas ( n = 6). Scale bar: 500 μm. ( D ) Representative images of H&E-stained aortic root sections with necrotic core outlined in dotted line ( n = 6 mice). Scale bar: 300 μm. ( E ) Representative images of aortic root sections stained against α-SMA (Cy3; red), MAC2 (Alexa Fluor 488 green), and cell nuclei (DAPI blue) with their subsequent quantification relative to lesion area ( n = 6). Scale bar: 300 μm. ( F ) Representative Picrosirius red staining of aortic root lesions with bars indicating fibrous cap thickness and quantification of fibrous cap thickness ( n = 6). Scale bar: 300 μm. ( G ) Schematic outlining experimental design and the generation of the EC transgenic ADAMTS7 mouse. Created with BioRender.com. ( H ) ORO staining of the en face aorta and quantification of lesion area. n = 3–4. ( I ) Representative images of H&E-stained aortic root sections and quantification of plaque areas ( n = 5–6). Scale bar: 500 μm. **** P < 0.0001, * P < 0.05. Statistics were analyzed using a 2-tailed Student’s t test.

Article Snippet: The sections then were stained using the following Abs and dilutions: 1:1,000 α-SMA-Cy3 (MilliporeSigma, C6198) and 1:500 MAC2 (Cedarlane, CL8942AP).

Techniques: Transgenic Assay, Staining

Journal: Neuroreport

Article Title: Galectin-3 expression in hippocampal CA2 following transient forebrain ischemia and its inhibition by hypothermia or antiapoptotic agents

doi: 10.1097/WNR.0000000000000538

Figure Lengend Snippet: Galectin-3 immunostaining in gerbil whole hippocampus including the CA2 sector at 60 h (upper) and 66 h (lower) after ischemic insult. In 60 h, weak immunoreactivity was recognized in CA2 (surrounded by arrowheads). In 66 h, prominent immunostaining was observed in CA2 (surrounded by arrowheads). Ependymal cells lining the lateral ventricles are strongly positive for galectin-3 as an internal positive control. The scale bar in the upper photograph is 500 μm.

Article Snippet: Anti-mouse galectin-3/Mac2 (Rat IgG, 14–5301) was purchased from Bay Bioscience Co. Ltd (Hyogo, Japan).

Techniques: Immunostaining, Positive Control

Journal: Neuroreport

Article Title: Galectin-3 expression in hippocampal CA2 following transient forebrain ischemia and its inhibition by hypothermia or antiapoptotic agents

doi: 10.1097/WNR.0000000000000538

Figure Lengend Snippet: H&E staining, immunohistochemical staining for galectin-3, Iba-1 and GFAP, and apoptotic DNA fragmentation detected by TUNEL staining in a low-power field and a high-power field of the hippocampus at 96 h after ischemic insult. Many damaged CA2 neurons had lost their nuclear affinity for hematoxylin and appeared as an eosinophilic ‘ghost neuron’ in H&E staining. The localization of galectin-3-positive cells coincided with that of Iba-1-positive cells. There were no GFAP-positive cells in the CA2 damaged area. Some of the TUNEL-positive CA2 neurons were distinguishable among the CA1 neurons by the shape of positive stained nuclei. Scale bars in LPF and HPF of H&E staining are 100 and 50 μm, respectively. G3, galectin-3; GF, GFAP; HE, HE staining; HPF, high-power field; Iba, Iba-1; LPF, low-power field; TU, TUNEL staining.

Article Snippet: Anti-mouse galectin-3/Mac2 (Rat IgG, 14–5301) was purchased from Bay Bioscience Co. Ltd (Hyogo, Japan).

Techniques: Staining, Immunohistochemical staining, TUNEL Assay

Journal: Neuroreport

Article Title: Galectin-3 expression in hippocampal CA2 following transient forebrain ischemia and its inhibition by hypothermia or antiapoptotic agents

doi: 10.1097/WNR.0000000000000538

Figure Lengend Snippet: Time course of representative microphotographs of immunohistochemical staining for galectin-3 in gerbil hippocampal CA2 sector at 0 (sham), 48, 60, and 96 h after ischemic insult. No Galectin-3 expression was detected in CA2 at 0 (sham) and 48 h. Microphotographs in the right column (HPF) show a high-power field of the rectangle on the left (LPF). Ependymal cells lining the lateral ventricles are strongly positive for galectin-3 as an internal positive control. Scale bars in LPF and HPF of sham are 200 and 50 μm, respectively. HPF, high-power field; LPF, low-power field.

Article Snippet: Anti-mouse galectin-3/Mac2 (Rat IgG, 14–5301) was purchased from Bay Bioscience Co. Ltd (Hyogo, Japan).

Techniques: Immunohistochemical staining, Staining, Expressing, Positive Control

Journal: Neuroreport

Article Title: Galectin-3 expression in hippocampal CA2 following transient forebrain ischemia and its inhibition by hypothermia or antiapoptotic agents

doi: 10.1097/WNR.0000000000000538

Figure Lengend Snippet: Intraischemic hypothermia and preadministration of TPCK and 2DG strongly inhibited the ischemia-induced galectin-3-positive cells in the CA2 region 96 h after ischemic insult. Data are presented as mean±SD. Numbers in parentheses indicate animal numbers in each group. *Statistically different from the ischemic control ( P <0.05), determined by ANOVA. ANOVA, analysis of variance; 2DG, 2-deoxy- d -glucose; ischemic cont., ischemic control; sham-op, sham operation; TPCK, N -tosyl- l -phenylalanyl chloromethyl ketone.

Article Snippet: Anti-mouse galectin-3/Mac2 (Rat IgG, 14–5301) was purchased from Bay Bioscience Co. Ltd (Hyogo, Japan).

Techniques: