Journal: Journal of Virology

Article Title: NFAT4 Is Required for JC Virus Infection of Glial Cells

doi: 10.1128/jvi.01456-06

Figure Lengend Snippet: FIG. 2. NFAT4 is functionally active in glial cells. (a) NFAT expression in SVG-A and U-87MG whole-cell lysates. NFATs 1 to 4 were detected using antibodies (NFATc1 7A6, NFATc2 4G6-G5, NFATc3 F-1, and NFATc4 H-74; Santa Cruz Biotech Inc.) diluted 1:200. Ramos cell lysates were used as a positive control. (b) Luciferase reporter gene assays using an NFAT-responsive reporter construct were used to compare NFAT activity in SVG-A cells and U-87MG cells in the presence or absence of 292 mg/liter glutamate. A control construct that lacked the NFAT binding site was used to measure basal transcriptional activity. (c) The NFAT reporter construct was cotransfected with either a control construct ( SV40 T-Ag) or a construct expressing the SV40 large T-Ag ( SV40 T-Ag).

Article Snippet: Ramos cell lysates were purchased from Santa Cruz Biotech Inc. Whole-cell extracts were separated on Tris-HCl-ready gels (Bio-Rad).

Techniques: Expressing, Positive Control, Luciferase, Construct, Activity Assay, Control, Binding Assay

Journal: Journal of Histochemistry & Cytochemistry

Article Title: Alveolar Epithelial Type I Cells Express β2-Adrenergic Receptors and G-protein Receptor Kinase 2

doi: 10.1369/jhc.3a6184.2004

Figure Lengend Snippet: Figure 3 Immunolocalization of GRK2 in whole lung sections of rat lung were double labeled with rabbit anti-GRK2 Ab (red) and goat anti- SP-C Ab (green), a marker for AT2 cells, and viewed using confocal mi- croscopy. (A) AECs are labeled with anti-GRK2 Ab with and without co- localization with anti-SP-C Ab. This sug- gests expression of GRK2 in both AT1 and AT2 cells. (B) Substitution of rab- bit IgG for anti-GRK2 Ab results in no specific staining. Original magnifi- cation 60. Immunolocalization of GRK2 in AEC in primary culture. (C) Cells labeled with anti-GRK2 Ab (green). (D) Cells labeled with normal rabbit IgG (green). Cell nuclei were identified using propidium iodide (red). AECs on day 5 that have trans- differentiated towards the AT1-like cell phenotype express immunoreac- tive GRK2 but are non-reactive with control IgG. Original magnification 40. Immunolocalization of GRK2 to isolated cells. (E,F) Freshly isolated AEC were double labeled with anti- GRK2 Ab (green) and VIIIB2 (red). IF images indicate that freshly isolated AT1 cells, reactive with the AT1 cell- specific marker VIIIB2, express GRK2 protein. A small cell (arrow) expresses GRK2 protein but does not react with VIIIB2. Original magnification 40.

Article Snippet: Commercially available control sample for GRK2 (Ramos cell lysate; Santa Cruz) was loaded with the AEC samples.

Techniques: Labeling, Marker, Expressing, Staining, Control, Isolation

Journal: Journal of Histochemistry & Cytochemistry

Article Title: Alveolar Epithelial Type I Cells Express β2-Adrenergic Receptors and G-protein Receptor Kinase 2

doi: 10.1369/jhc.3a6184.2004

Figure Lengend Snippet: Figure 4 Expression of GRK2 protein in AECs. Western analysis was performed with anti-GRK2 Ab using lysates of either purified freshly isolated AT2 cells (d.0) or AEC in culture that have transdif- ferentiated to the AT1 cell phenotype (days 6–8). (A) Representa- tive Western blot demonstrates similar amounts of GRK2 protein in AT2 and AT1-like cells in culture. (B) Summary of densitometry from Western blotting analyses (n3). Bar graph shows average densitometric values ( SEM) normalized to GRK2 protein abun- dance on d.0 (100%). There was no significant difference in GRK2 expression between d.0 and d.6–8 (p0.58).

Article Snippet: Commercially available control sample for GRK2 (Ramos cell lysate; Santa Cruz) was loaded with the AEC samples.

Techniques: Expressing, Western Blot, Purification, Isolation

Journal: Biology of reproduction

Article Title: Members of the Toll-like receptor family of innate immunity pattern-recognition receptors are abundant in the male rat reproductive tract.

doi: 10.1095/biolreprod.106.059410

Figure Lengend Snippet: FIG. 2. Immunoblot analysis of TLR proteins in male reproductive organs. Aliquots (100 lg) of cytoplasmic protein extracts were separated by PAGE, electroblotted, and blots probed with anti-TLR antibodies followed by enhanced chemiluminescence detection using Pierce Super- Signal West Pico or Pierce SuperSignal West Femto (*F) substrate. Representative results are shown (n ¼ 3–5 rats). þ, control rat tissue extracts from spleen, TLRs 1–7; lung, TLR8; and small intestine, TLR9. þ Ext, positive-control whole cell lysates used were Raw 264 Abelson transformed macrophages (TLRs 1, 2–6, and 8–10); SW480 colorectal adenocarcinoma (TLR2); Daudi cell extract (TLR7); Ramos cell lysate (TLR10) and mouse heart whole cell lysate (TLR11). Blots were stripped and reprobed with anti-actin monoclonal antibody to detect actin as a loading control. Representative results are shown (n ¼ 3–5 rats) for blots exposed to film for the same length of time when using equivalent chemiluminescent substrate.

Article Snippet: Mouse heart tissue lysate (40102) and Ramos cell lysate (40175) was from Imgenex.

Techniques: Western Blot, Control, Positive Control, Transformation Assay

Journal: Nano-Micro Letters

Article Title: Multiplexed Profiling of Extracellular Vesicles for Biomarker Development

doi: 10.1007/s40820-021-00753-w

Figure Lengend Snippet: Summary of the external coding-based multiplexing profiling of EV components

Article Snippet: Aptamer/AuNPs based coding , Colorimetry , Surface proteins: CD63, EpCAM, PDGF, PSMA, PTK7 , CCM from HeLa, PC-3, Ramos, CEM cell lines , Ultracentrifugation , LoD = 3.2 μg mL −1 , Low production of PSMA in PC‐3 cells, higher level of PTK7 from CEM exosomes than that of Ramos exosomes, absence or very low presence of PDGF in all exosomes , [ ] .

Techniques: Multiplexing, Isolation, Colorimetric Assay, Polymer, Fluorescence, Clinical Proteomics, Expressing, Biomarker Discovery, Derivative Assay, Ab Array, Marker, Extraction

Journal: Nucleic Acids Research

Article Title: Programmable manipulation of oligonucleotide–albumin interaction for elongated circulation time

doi: 10.1093/nar/gkac156

Figure Lengend Snippet: In vitro test on the specificity and stability of Sgc8. ( A ) Binding ability to HCT116 cells and Ramos cells tested with flow cytometric assay. Unmodified Sgc8 was used as the positive control, and library (lib) was used as the negative control. The cells were incubated with DNAs respectively at a concentration of 250 nM in a binding buffer for 30 min. The incubation temperature was 4°C. ( B ) Stability of Sgc8 variant after incubation with 10% FBS for 48 h analysis by PAGE. ( C ) Flow cytometric assay of HCT116 cells treated with Sgc8 and Sgc8-F23 after incubation with 10% FBS for 0–36 h, illustrating the binding ability after the stability test.

Article Snippet: Human colon cancer cell line HCT116 was purchased from ATCC, Human leukemia cell Ramos was purchased from China Center for Type Culture Collection (CCTCC, China).

Techniques: In Vitro, Binding Assay, Flow Cytometry, Positive Control, Negative Control, Incubation, Concentration Assay, Variant Assay

Journal: Nucleic Acids Research

Article Title: Programmable manipulation of oligonucleotide–albumin interaction for elongated circulation time

doi: 10.1093/nar/gkac156

Figure Lengend Snippet: The binding affinity of Sgc8 derivatives to albumin. ( A ) Binding affinity analysis of Sgc8 derivatives and Sgc8 with aldehyde/sulfate latex beads coated HSA at room temperature. ( B ) HSA was immobilized to 3.8 μm 40 mg/ml aldehyde/sulfate latex beads (Thermo Fisher Scientific, Waltham, MA USA), after incubation with sgc8 or Sgc8-F23, the beads were washed to remove excess aptamers. The fluorescence signal of HCT116 and Ramos cells treated with the beads (sgc8 or Sgc8-F23 binding with HSA) was collected by flow cytometer.

Article Snippet: Human colon cancer cell line HCT116 was purchased from ATCC, Human leukemia cell Ramos was purchased from China Center for Type Culture Collection (CCTCC, China).

Techniques: Binding Assay, Incubation, Fluorescence, Flow Cytometry