Journal: Pathophysiology

Article Title: Metabolic Syndrome-Driven Changes in Cardiac Lymphatic Endothelium: mRNA Expression and Emerging Questions

doi: 10.3390/pathophysiology33010004

Figure Lengend Snippet: RT-PCR analysis. Selected mRNA expression in sorted LECs from db/db and control mice hearts. Some mRNA levels significantly increase in db/db mice compared to controls. Statistically significant differences ( p -value < 0.05) are marked (*). Abbreviations: Acta2—Actin 2/smooth muscle actin; Bax—BCL2 Associated X; Bcl2—B-cell CLL/lymphoma 2; CCL21—Chemokine (C-C motif) ligand 21; CEACAM1—Carcinoembryonic antigen-related cell adhesion molecule 1; CPT1A—Carnitine Palmitoyltransferase 1A; CX3CL1—chemokine (C-X3-C motif) ligand 1; eNOS—endothelial nitric oxide synthase;Fbn1—fibrillin-1; FOXO1—Forkhead Box O1; GATA2—GATA-binding factor 2; ICAM-1—intercellular adhesion molecule 1; KLF2—Krüppel-like Factor 2; MMP2—Metalloproteinase 2; PDPN—podoplanin; PTEN—phosphatase and tensin homolog deleted on chromosome ten; Reln—reelin; Sdc4—syndecan 4; Snail1—Snail family transcriptional repressor-1; Snail2—Snail family transcriptional repressor-2; VCAM-1—vascular cell adhesion molecule 1; VE-cadherin—Vascular Endothelial Cadherin.

Article Snippet: To detect mRNA levels of selected genes, the following TaqMan Gene Expression Assays were used: CD36: Mm00432403_m1; Pten: Mm00477208_m1; Foxo1: Mm00490671_m1; Klf2: Mm00500486_g1; Bax: Mm00432051_m1; Bcl2: Mm00477631_m1; Cpt1a: Mm01231183_m1; Ccl21a: Mm03646971_gH; Cx3cl1: Mm00436454_m1; Icam1: Mm00516023_m1; Vcam1: Mm01320970_m1; Emilin1: Mm00467244_m1; Mmp2: Mm00439498_m1; Reln: Mm00465200_m1; Snai1: Mm00441533_g1; Snai2: Mm00441531_m1; Acta2: Mm00725412_s1; Cdh5: Mm00546194_s1; Gata2: Mm00492301_m1; Cldn5: Mm00727012_s1; Ceacam1: Mm04204476_m1; Pdpn: Mm01348912_g1; Fbn1: Mm00435217_m1; Nos3: Mm00435217_m1; Sdc4: Mm00488527_m1.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Binding Assay

Journal: Scientific Reports

Article Title: Spatial mapping of influenza and coronavirus receptors in the respiratory and intestinal tract epithelium of beef cattle using advanced PixF image analysis

doi: 10.1038/s41598-025-28429-0

Figure Lengend Snippet: Coronavirus receptors in the respiratory tract and intestinal tracts of beef cattle. Confocal images showing angiotensin-converting enzyme 2 (ACE2, A ), transmembrane protease serine 2 (TMPRSS2, B ), aminopeptidase N (APN, C ), dipeptidyl peptidase 4 (DPP4, D ), and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1, E ) expression following immunofluorescence staining. Primary antibodies, ACE2 (4 µg/ml, Cat# sc-390851), TMPRSS2 (0.2 µg/ml; Cat# sc-515727), APN (0.16 µg/ml; Cat# sc-166105), CEACAM1 (4 µg/ml; Cat# sc-166453), and DPP4 (1:50 dilution; Cat# BOV2078, Washington State University Monoclonal Antibody Center, Pullman, WA, USA); Secondary antibody, AffiniPure™ Donkey Anti-Mouse IgG (H + L) conjugated Alexa Fluor ® 488 (15 µg/ml; Cat# 715-545-150). Blue represents nuclear staining using DAPI; scale bar = 20 µM. Representative images of coronavirus receptor expression across tissues are shown. The epithelial lining of the trachea ( Ai ), alveoli ( Aiv ), small intestine ( Av ), and large intestine ( Avi ) showed strong ACE2 expression compared to the bronchus ( Aii ) and bronchiole ( Aiii ). The respiratory epithelium showed extensive TMPRSS2, APN, and DPP4 expression with a multifocal expression on the alveoli ( Bi – iv , Ci – iv , Di – iv ). TMPRSS2 ( Bv – vi ) and DPP4 ( Dv – vi ) were detected uniformly on villi and crypt regions in the small and large intestine epithelia, while APN levels were comparatively low in both intestines ( Cv – vi ). With the exception of the bronchus ( Eii ) and bronchiole ( Eiii ), the levels of CEACAM1 were low to no detection on the epithelia of all the tissues assessed ( Ei , Eiv – vi ).

Article Snippet: Deparaffinized and heat-treated antigen retrieval sections were incubated with Animal-Free Blocker R.T.U (Cat# SP-5035, Vector Laboratories) for 30 min at room temperature, followed by incubation with optimized concentrations of primary antibodies: ACE2 (4 μg/ml, Cat# sc-390851, Santa Cruz Biotechnology, Dallas, TX, USA), TMPRSS2 (0.2 μg/ml; Cat# sc-515727, Santa Cruz Biotechnology), APN (0.16 μg/ml; Cat# sc-166105, Santa Cruz Biotechnology), CEACAM1 (4 μg/ml; Cat# sc-166453, Santa Cruz Biotechnology), and DPP4 (1:50 dilution; Cat# BOV2078, Washington State University Monoclonal Antibody Center, Pullman, WA, USA) overnight (16 h) at 4 °C.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Spatial mapping of influenza and coronavirus receptors in the respiratory and intestinal tract epithelium of beef cattle using advanced PixF image analysis

doi: 10.1038/s41598-025-28429-0

Figure Lengend Snippet: Heat map showing the distribution of influenza and coronavirus receptors in the respiratory and intestinal tracts of beef cattle. Heatmap was generated using data from PixF image analysis showing the average log 10 total intensity (sum of intensity in all the pixels with signal) of cropped images ( n = 3) by a 350*350-pixel region of interest (ROI) on the epithelium of respiratory and intestinal tract tissues. The color scale of lectin staining (orange, high; light yellow, medium; green, low) and immunofluorescence assay (red, high; white, medium; blue, low; dark blue < 5) indicates the level of log total intensity. ACE2, angiotensin-converting enzyme 2; TMPRSS2, transmembrane protease serine 2; APN, aminopeptidase N; DPP4, dipeptidyl peptidase 4; CEACAM1, carcinoembryonic antigen-related cell adhesion molecule 1. Respiratory and intestinal tissues were analyzed separately. For lectin staining data was analyzed with two-way ANOVA, and multiple comparisons with post hoc Tukey test. The levels of sialic acids within the same tissue ( + denotes significant higher with P < 0.05) and one SA across different tissues ( # denotes significant higher with P < 0.05). Immunofluorescence assay was analyzed with one-way ANOVA (within the same receptor), and multiple comparison with post hoc Tukey test was conducted to compare between different tissues ( * denotes significant higher with P < 0.05).

Article Snippet: Deparaffinized and heat-treated antigen retrieval sections were incubated with Animal-Free Blocker R.T.U (Cat# SP-5035, Vector Laboratories) for 30 min at room temperature, followed by incubation with optimized concentrations of primary antibodies: ACE2 (4 μg/ml, Cat# sc-390851, Santa Cruz Biotechnology, Dallas, TX, USA), TMPRSS2 (0.2 μg/ml; Cat# sc-515727, Santa Cruz Biotechnology), APN (0.16 μg/ml; Cat# sc-166105, Santa Cruz Biotechnology), CEACAM1 (4 μg/ml; Cat# sc-166453, Santa Cruz Biotechnology), and DPP4 (1:50 dilution; Cat# BOV2078, Washington State University Monoclonal Antibody Center, Pullman, WA, USA) overnight (16 h) at 4 °C.

Techniques: Generated, Staining, Immunofluorescence, Comparison