Journal: Cell reports

Article Title: MicroRNA mediated regulation of the onset of enteroblast differentiation in the Drosophila adult intestine

doi: 10.1016/j.celrep.2022.111495

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-V5 , Bio-Rad , Cat# MCA1360GA; RRID:AB_567249.

Techniques: Recombinant, Electron Microscopy, Plasmid Preparation, Protease Inhibitor, SYBR Green Assay, Western Blot, Blocking Assay, In Situ, Software, Real-time Polymerase Chain Reaction, Microscopy

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Enolase 1 suppresses influenza A virus replication by blocking the nuclear import of the viral ribonucleoprotein complex

doi: 10.1007/s00018-026-06146-9

Figure Lengend Snippet: ENO1 interacts with the NP protein. ( A , B ) HEK293T cells were transfected individually or in combination with Flag-tagged ENO1 and HA-tagged NP. At 24 h post-transfection, lysates of cells were immunoprecipitated with mouse anti-Flag mAb (A) or mouse anti-HA mAb (B) and then western blotted with rabbit anti-Flag pAb and rabbit anti-HA pAb, respectively. ( C ) HEK293T cells were infected with H1N1 virus (MOI = 5). At 12 h p.i., lysates from HEK293T cells were immunoprecipitated with mouse anti-ENO1 pAb, followed by western blotting with rabbit anti-ENO1 pAb and rabbit anti-NP pAb, respectively. ( D ) Purified GST and GST-ENO1 protein were used to pull down Flag-tagged NP. ( E ) A549 cells were uninfected or infected with H1N1 virus (MOI = 5). At 12 h p.i., the cells were fixed and stained with mouse anti-NP mAb and rabbit anti-ENO1 pAb. Scale bars, 5 μm. ( F , G ) HEK293T cells were transfected with the indicated plasmids. At 24 h post-transfection, cell lysates were immunoprecipitated with mouse anti-Flag mAb, followed by western blotting with rabbit anti-Flag pAb, rabbit anti-HA pAb, and rabbit anti-V5 pAb. The data are representative of at least three independent experiments

Article Snippet: The following primary antibodies were obtained from commercial sources: mouse anti-Flag mAb (A00187 -100, GenScript), rabbit anti-Flag pAb (20543-1-AP, Proteintech) mouse anti-HA mAb (A01244-100, GenScript), rabbit anti-HA pAb (51064-2-AP, Proteintech), mouse anti-V5 mAb (A01724-100, GenScript), rabbit anti-V5 pAb (14440-1-AP, Proteintech), rabbit anti-GAPDH pAb (10494-1-AP, Proteintech), rabbit anti-ENO1 pAb (11204-1-AP, Proteintech), rabbit anti-LaminA/C pAb (10298-1-AP, Proteintech), rabbit anti-KPNA4 pAb (12463-1-AP, Proteintech), rabbit anti-PB2 pAb (GTX125926, GeneTex), rabbit anti-PB1 pAb (GTX125923, GeneTex), rabbit anti-PA pAb (GTX118991, GeneTex), rabbit anti-NP pAb (GTX125989, GeneTex).

Techniques: Transfection, Immunoprecipitation, Western Blot, Infection, Virus, Purification, Staining

Journal: Cell reports

Article Title: Global Analysis of Intercellular Homeodomain Protein Transfer

doi: 10.1016/j.celrep.2019.06.056

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal anti-V5 antibody , GenWay Biotech , RRID: AB_10280227.

Techniques: Recombinant, In Vitro, Transfection, Protease Inhibitor, Control, Plasmid Preparation, Software, Expressing, Fluorescence

Journal: Journal of Lipid Research

Article Title: Diet1, bile acid diarrhea, and FGF15/19: mouse model and human genetic variants [S]

doi: 10.1194/jlr.M078279

Figure Lengend Snippet: DIET1 variant rs12256835 influences secretion of FGF19 in a cell-based assay. A: Representative results of an FGF19 secretion assay comparing Diet1-1721H and -1721Q. FGF19-Myc was expressed in HEK293T cells in combination with either Diet1-1721H-V5, Diet1-1721Q-V5, or an irrelevant control protein (Kdm5c-V5). Cellular Diet1 protein levels and secreted FGF19 levels were visualized by Western blot. MW, molecular mass in kD. B: Quantification of FGF19 secretion from nine experiments of the type shown in A. Each point is one independent transfection (done in triplicate or quadruplicate), representing the amount of FGF19 secreted per unit of cellular V5 expression for control protein (Kdm5c), Diet1-1721H, or Diet1-1721Q. The overall P-value for all three conditions is P = 3.70 × 10−7 (one-way ANOVA). Individual comparisons were made using Tukey’s HSD post hoc test on log2-normalized values. Small horizontal lines represent geometric mean of samples for each protein isoform (*P < 0.05, **P < 0.01). C: Representative results of an FGF19 secretion assay comparing Diet1-1721D and -1721G. Details are as in A. D: Quantification of FGF19 secretion from five experiments of the type shown in C. No statistically significant difference (n.s.) in the amount of extracellular FGF19 was detected between Diet1-1712D and -1712G (P = 0.945; one-way ANOVA).

Article Snippet: Diet1 was detected with rabbit anti-V5 (1:5,000; Bethyl Laboratories, Montgomery, TX); FGF19 was detected with rabbit anti-Myc (1:10,000; Bethyl Laboratories); and β-actin was detected with a mouse monoclonal antibody (1:10,000; Sigma, St. Louis, MO).

Techniques: Variant Assay, Cell Based Assay, Western Blot, Transfection, Expressing

Journal: Journal of Lipid Research

Article Title: Diet1, bile acid diarrhea, and FGF15/19: mouse model and human genetic variants [S]

doi: 10.1194/jlr.M078279

Figure Lengend Snippet: Diet1-1721H and -1721Q isoforms do not differentially affect Diet1-FGF19 interaction. Coimmunoprecipitation of Diet1-1721H and Diet1-1721Q with FGF19. FGF19-Myc was coexpressed in HEK293T cells with Diet1-1721H-V5 or Diet1-1721Q-V5. Input lanes show similar levels of expression of the two Diet1 isoforms. Bands corresponding to precipitation of FGF19 with Diet1-1721H are on the left of the dashed line, and bands corresponding to precipitation of FGF19 with Diet1-1721Q are on the right of the dashed line. Dashed boxes indicate immunoprecipitation of Diet1 (V5 IP); solid boxes indicate immunoprecipitation of FGF19 (Myc IP). Both Diet1 isoforms interact with FGF19 with similar efficiency. A representative blot from eight experiments is shown.

Article Snippet: Diet1 was detected with rabbit anti-V5 (1:5,000; Bethyl Laboratories, Montgomery, TX); FGF19 was detected with rabbit anti-Myc (1:10,000; Bethyl Laboratories); and β-actin was detected with a mouse monoclonal antibody (1:10,000; Sigma, St. Louis, MO).

Techniques: Expressing, Immunoprecipitation