Journal: World Journal of Diabetes

Article Title: Linagliptin alleviates fatty liver disease in diabetic db / db mice

doi: 10.4239/wjd.v7.i19.534

Figure Lengend Snippet: Immunohistochemical staining for lymphatic vessel endothelial hyaluronan receptor-1 in the liver of intact (A and B), placebo-treated (C) and linagliptin-treated (D, E and F) db / db mice. Staining by anti-LYVE-1 antibodies, indirect streptavidin-biotin method; A, C, E and F: × 400; B and D: × 100. LYVE-1: Lymphatic vessel endothelial hyaluronan receptor-1.

Article Snippet: Blocking of endogenous peroxidase was performed by incubating the sections in a 0.3% H 2 O 2 solution for 10 min with a subsequent incubation in normal horse non-immune blocking serum for 20 min. Next, the sections were incubated for one hour at room temperature with anti-LYVE-1 (Isotype: Rabbit polyclonal, bs-1311R; Bioss) at a final dilution of 5 mg/mL; washed in 3 changes of phosphate buffer for 3 min; and further incubated for 30 min at room temperature with a biotinylated second antibody followed by washing in 3 changes of phosphate buffer for 5 min. Incubation with the ABC-peroxidase complex was performed for 30 min at room temperature followed by washing in 3 changes of phosphate buffer for 5 min. Immunohistochemical staining of the sections was performed with a chromogenic substrate (ImmPACT DAB, Vector Laboratories, United States).

Techniques: Immunohistochemical staining, Staining

Journal: bioRxiv

Article Title: Multi-omics Analysis Identifies IgG2b Class-Switching with ALCAM-CD6 Co-Stimulation in Lymph Nodes During Advanced Inflammatory-Erosive Arthritis

doi: 10.1101/2022.10.27.514103

Figure Lengend Snippet: PLNs from WT (5.5-month-old; n=10 PLNs) (A) and TNF-Tg mice with Early (5-6-month-old; n=6 PLNs) (B) and Advanced (>8-month-old; n=12 PLNs, 6/capture area) (C-D) arthritis were harvested and processed for spatial transcriptomics. Each H&E-stained image (left) corresponds to a transcriptional representation (right) of the PLN sinus (red spots overlaying white sinus spaces), paracortical (green spots), and follicular (blue spots) regions. Isolated spots with high ribosomal gene content were annotated as an undefined (purple) cell population (A-E) . Feature plots represent the identification of Lyve Hi /Marco Hi sinuses (F-G) , Ptprc Hi paracortices (H) , and Ptprc Hi /Ms4a1 Hi follicles (I) that were validated by histologic localization as in A-D . Unsupervised clustering (resolution=0.5) resolved 11 transcriptionally distinct PLN regions, where the sinus-associated spots localized to 3 distinct clusters (dashed lines) (J) . The Ighg1 Hi / Ighg2b Hi / Ighg2c Hi / Ighm Hi Sinus-Immuno (K-N) and Lyve1 Hi / Marco Hi Sinus-LEC (F-G) populations predominated, while Sinus-Undefined spots exhibited a high proportion of ribosomal genes. The predominant Sinus-Immuno (red) and Sinus-LEC (blue) populations were subset and re-clustered (O) , and individual PLNs were evaluated for total sinus-associated spots (Sinus-Immuno + Sinus-LEC) (P) and proportion of Sinus-Immuno spots (Q) . Statistics: One-way ANOVA, *p<0 . 05, **p<0 . 01 (P , Q) .

Article Snippet: Antibodies and dilutions for immunofluorescent staining: Rabbit anti-mouse CD6 (ThermoFisher, Cat# MA5-29680, RRID: AB_2785505, 1:50), rat anti-mouse F4/80 (BioRad, Cat# MCA497R, RRID:AB_323279, 1:50), rabbit anti-mouse Marco (Abcam, Cat# ab239369, 1:100), Alexa Fluor 555 goat anti-mouse IgG2b cross-adsorbed (ThermoFisher Scientific, Cat# A-21147, RRID:AB_2535783, 1:50), APC rat anti-mouse CD45R/B220 (BD Biosciences, Cat# 553092, RRID:AB_398531, 1:100), goat anti-mouse Alcam (CD166; ThermoFisher, Cat# PA5-47083, RRID:AB_2607383, 1:100), Cy3 goat anti-mouse IgM (Jackson ImmunoResearch Laboratories, Cat# 115-165-020, RRID:AB_2338683, 1:200), FITC donkey anti-mouse IgG (Jackson ImmunoResearch Laboratories, Cat# 715-095-150, RRID:AB_2340792, 1:200), goat anti-proliferating cell nuclear antigen (PCNA; Santa Cruz Biotechnology, Cat# sc-9857, RRID:AB_2160372, 1:50), FITC Peanut Agglutinin (PNA; Millipore Sigma, L7381-1MG, 1:50), goat anti-mouse peripheral node addressin (PNAd; BD Biosciences, Cat#553863, RRID:AB_395099, 1:50), rabbit anti-mouse Lyve1 (Acris Antibodies, Cat# DP3513, RRID:AB_1004776, 1:50), AlexaFluor 568 donkey anti-goat IgG (ThermoFisher Scientific, Cat# A-11057, RRID:AB_142581, 1:200), AlexaFluor 647 F(ab’) 2 fragment donkey anti-rat IgG (Jackson ImmunoResearch, Cat# 712-606-153, RRID:AB_2340696, 1:200), and AlexaFluor 647 donkey anti-rabbit IgG (Abcam, Cat# ab150075, RRID:AB_2752244, 1:400).

Techniques: Staining, Isolation

Journal: Current Oncology

Article Title: Characteristic of Uterine Rhabdomyosarcoma by Algorithm of Potential Biomarkers for Uterine Mesenchymal Tumor

doi: 10.3390/curroncol29040190

Figure Lengend Snippet: Significance of lymphatic endothelial cells in the primary Case 1 uterine tumor. Tumor cells infiltrating the lymph vessels within the uterine tumor are round cells with a high nuclear cytoplasmic ratio (N/C) (upper left panel). Circular cells with a high nuclear cytoplasm ratio (N/C) were also found in the biopsy from the tumor of the left supraclavicular lymph node (upper right panel). The results indicate that tumor cells from the primary uterine tumor formed metastases in the left supraclavicular lymph node by lymphatic metastasis. H&E staining and immunofluorescence of uterine leiomyosarcoma and the Case 1 uterine tumor with an anti-human CD31 (green) and anti-human LYVE-1 antibody (red) (lower panels). Human vascular endothelial cells and lymphatic endothelial cells were detected as CD31 and LYVE-1 double-positive cells. The tumor cells from the Case 1 uterine tumor appear to differentiate into lymphatic endothelial cell progenitors (lower light panel), but the tumor cells from the uterine leiomyosarcoma do not appear to have differentiated to lymphatic endothelial cell progenitors (lower left panel).

Article Snippet: The antibody for LYVE-1 (bs-20353R) was purchased from Bioss Inc. (Boston, MA, USA).

Techniques: Staining, Immunofluorescence