Journal: Viruses

Article Title: Recombinant Pseudorabies Virus with TK/gE Gene Deletion and Flt3L Co-Expression Enhances the Innate and Adaptive Immune Response via Activating Dendritic Cells

doi: 10.3390/v13040691

Figure Lengend Snippet: Characterization of the recombinant HNX-TK − /gE − -Flt3L. ( A ) Detection of the transcription of the Flt3L gene in HNX-TK − /gE − -Flt3L virus- and HNX-infected cells through RT-qPCR (**** < 0.001). ( B ) Detection of the Flt3L protein and viral gB protein expression through an immunofluorescence assay (IFA). Green: gB-positive cells; red: Flt3L-positive cells; blue: DAPI-stained PK-15 cell nucleus. DAPI, 4’,6-diamidino-2-phenylindole. ( C ) Western blot analysis of β-actin, gD, and Flt3L of the PK-15 cells infected with HNX-TK − /gE − -Flt3L, HNX, or DMEM as a control. ( D ) One step growth curves of HNX-TK − /gE − -Flt3L and HNX in PK-15 cells.

Article Snippet: The blots were blocked by skim milk (BD, Bioscience, Inc., Saint Louis, MO, USA), and this was followed by incubation with rabbit anti-Flt3L polyclonal antibody (Bioss, Beijing, China), mouse anti-gD monoclonal antibody (Keqian Ltd., Wuhan, China), and mouse anti-β-actin monoclonal antibody (Bioss).

Techniques: Recombinant, Infection, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Western Blot

Journal: Oncology Letters

Article Title: Comparison of the expression of TGF-β1, E-cadherin, N-cadherin, TP53, RB1CC1 and HIF-1α in oral squamous cell carcinoma and lymph node metastases of humans and mice

doi: 10.3892/ol.2017.7456

Figure Lengend Snippet: Expression of tumor protein (TP)53 at different stages in the progression of oral carcinoma in human and mouse samples. (A) Photomicrographs of immunohistochemical positive expression of TP53 (scoring 2/3) in normal and dysplastic oral mucosa, OSCC and submandibular lymph node in lymphatic nodes metastatic carcinoma (magnification, ×200). (B and C) Data relative to the expression of TP53 at the different cancer stages. (D) Positive expression rate of TP53 at different cancer stages in human and mouse samples. *Compared with normal stage in human samples, P<0.05; ∆compared with normal or dysplasia stages in human samples, P<0.05; #compared with normal or dysplasia stages in mouse samples, P<0.05.

Article Snippet: The antibodies used in the present study were as follows: Mouse monoclonal antibodies for E-cadherin and p53 (ZSGB-Bio, China), rabbit polyclonal antibodies for TGF-β1 and HIF-1α (Boster Biological Technology, China), mouse monoclonal antibody for N-cadherin (Santa Cruz, USA), and rabbit polyclonal antibodies for RB1CC1 (Proteintech Group, China).

Techniques: Expressing, Immunohistochemical staining

Journal: PLoS ONE

Article Title: In Vivo Serial MR Imaging of Magnetically Labeled Endothelial Progenitor Cells Homing to the Endothelium Injured Artery in Mice

doi: 10.1371/journal.pone.0020790

Figure Lengend Snippet: 7 days after in vitro differentiation, spleen-derived EPCs showed expression of CD31 (B), CD34 (C) and vWF (D) with the cytoplasm stained in brown. Photo A is the control. Hematoxylin-stained nuclei were shown in blue. (Magnification ×400).

Article Snippet: After 7 days in culture, spleen-derived EPCs in 4 wells of a 24-well culture plate were extensively washed, and fixed in ice-cold 4% paraformaldehyde for 20 min. Immunocytochemistry was performed using antibodies against CD31, CD34 (Wuhan Boster Biological Technology, LTD. Wuhan, China) and von Willebrand factor (vWF) ( Zhongshan Goldenbridge Biotechnology Co., LTD. Beijing, China) according to standard protocols.

Techniques: In Vitro, Derivative Assay, Expressing, Staining

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Summary of the immunocytochemical analysis of CNHCs with antibodies against CD45, CD31, and CAIX

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (−UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX (A) . Clusters of CNHC-UMF and -BF without reactivity for CAIX antibodies ( B and C , respectively). A single CNHC-MF with positive cytoplasmic (D) and without staining for CAIX (E) . Single CAIX-negative CNHC-UMF and -BF ( F and G , respectively).

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques: Marker, Staining

Journal: Cell Reports Medicine

Article Title: Truncating NFKB1 variants cause combined NLRP3 inflammasome activation and type I interferon signaling and predispose to necrotizing fasciitis

doi: 10.1016/j.xcrm.2024.101503

Figure Lengend Snippet: Truncating variants of NFKB1 impair autophagy (A) MitoSOX staining of untreated NFKB1 R157∗/R157∗ and WT THP-1 monocytes was analyzed by flow cytometry. Quantifications and representative plots are shown. (B) HMDMs (left) or NFKB1 R157∗/R157∗ and WT THP-1 monocytes (right) were activated with LPS, and MitoTempo was applied 1 h before ATP. Secreted mature IL-1β was detected by ELISA. (C) ρ 0 or conventional NFKB1 R157∗/R157∗ and WT THP-1 monocytes were activated with LPS and secreted mature IL-1β was detected by ELISA. (D) HMDMs (left) or NFKB1 R157∗/R157∗ and WT THP-1 monocytes (right) were activated with LPS for indicated times and SQSTM1 was blotted from lysates. Quantifications, representative blots, and TPL are shown. (E) NFKB1 R157∗/R157∗ and WT THP-1 monocytes were inhibited with E-64d and pepstatin A (abbreviated as EP) 1 h prior to LPS (for indicated times). LC3 was blotted from cell lysates. Quantifications, representative blot, and TPL are shown. (F and G) NFKB1 R157∗/R157∗ and WT THP-1 monocytes were treated with LPS and (F) 3-MA (6 h 45 min) or (G) rapamycin (17 h 45 min) and ATP (for the last 45 min). Secreted mature IL-1β was detected by ELISA and is expressed as fold change relative to the respective cells stimulated with LPS and ATP. (H) NFKB1 R157∗/R157∗ and WT THP-1 monocytes were activated with LPS (18 h). Cells staining positive for Hoechst (nuclear stain) and autophagic vesicle marker were analyzed by flow cytometry. Quantitation and representative plots are shown. (I) Schematic representation of the effects of truncating NFKB1 mutations on macroautophagy and NLRP3 inflammasome activation. Wilcoxon test (A), one-way RM-ANOVA (B), or two-way ANOVA (C–H) followed by Dunnett’s (B), Šidak’s (D–G), or Tukey’s test (C and H). The data are shown as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (A) n = 8; (B) HMDMs: 1 control, 3 variant carriers, THP-1 n = 4; (C–E) THP-1 n = 4; (D) HMDMs: 2 controls, 4 variant carriers; (F–H) THP-1 n = 4. See also Figure S4 .

Article Snippet: For staining with LC3 antibody (Novus Biologicals NB100-2331) THP-1 monocytes were centrifuged and lyzed in sucrose lysis buffer (250 mM Sucrose, 1 mM EDTA, and 1x protease inhibitor).

Techniques: Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Marker, Quantitation Assay, Activation Assay, Control, Variant Assay

Journal: Cell Reports Medicine

Article Title: Truncating NFKB1 variants cause combined NLRP3 inflammasome activation and type I interferon signaling and predispose to necrotizing fasciitis

doi: 10.1016/j.xcrm.2024.101503

Figure Lengend Snippet:

Article Snippet: For staining with LC3 antibody (Novus Biologicals NB100-2331) THP-1 monocytes were centrifuged and lyzed in sucrose lysis buffer (250 mM Sucrose, 1 mM EDTA, and 1x protease inhibitor).

Techniques: Control, Recombinant, Lysis, Protease Inhibitor, Western Blot, Cell Culture, Molecular Weight, Staining, Sequencing, Modification, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, Transfection, Quantitation Assay, Mutagenesis, Software, CRISPR, Variant Assay, Cytometry