Journal: PloS one

Article Title: Identification of PBX1 target genes in cancer cells by global mapping of PBX1 binding sites.

doi: 10.1371/journal.pone.0036054

Figure Lengend Snippet: Figure 6. MEOX1 interacts with PBX1 and mediates its growth effect. A. HEK293 cells were transfected with PBX1-V5 and/or MEOX1-FLAG expression vectors. Immunoprecipitation and Western blot were performed using epitope tag-specific antibodies. B. Left panel: OVCAR3 cells were transfected with MEOX1 expression vector or control vector. Western blot was performed to test MEOX1 expression (top). The same cells were transfected with PBX1 siRNA or control siRNA and Western blot was performed to test the down-regulation of PBX1 protein (middle). Detection of GAPDH protein was used as a loading control (bottom). Right panel: Relative cell numbers were measured in OVCAR3 cells transfected with MEOX1 cDNA, PBX1 siRNA, control plasmid (pLPC), and control siRNA (siLuc). Student’s t-test was used to determine the significance between the MEOX1 over-expressed group and the control group. doi:10.1371/journal.pone.0036054.g006

Article Snippet: Western blot was performed using an anti-FLAG antibody (Sigma, St. Louis, MO) to detect MEOX1 protein or an anti-V5 antibody (Bethyl, Montgomery, TX) to detect PBX1 protein.

Techniques: Transfection, Expressing, Immunoprecipitation, Western Blot, Plasmid Preparation, Control

Journal: iScience

Article Title: tANCHOR-cell-based assay for monitoring of SARS-CoV-2 neutralizing antibodies rapidly adaptive to various receptor-binding domains

doi: 10.1016/j.isci.2024.109123

Figure Lengend Snippet:

Article Snippet: Rabbit anti-V5 , Novus Biologicals , Cat# NB600-381.

Techniques: Virus, Recombinant, Saline, Staining, Isolation, Protease Inhibitor, Software, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: European journal of immunology

Article Title: A common human TLR1 polymorphism regulates the innate immune response to lipopeptides.

doi: 10.1002/eji.200737034

Figure Lengend Snippet: Figure 2. Immunofluorescence and expression studies of TLR1 variants. (A) TLR1 coding region and SNP positions; LRR, leucine-rich region; TM, transmembrane domain. SNP N248S and I602S are marked with a circle. (B–F) Localization patterns of V5-epitope-tagged TLR1 variants that were transfected into HEK293 cells and visualized by immunofluorescence. The TLR1 constructs varied at amino acids 248 (N or S) and 602 (I or S) and include TLR1_NI (248S_602I) (B), NS (248N_602S) (C), SS (248S_602S) (D), SI (248S_602I) (E), and a pEF6 empty vector control (F). (G) HEK293 cells were transfected with control vector (pEF6 without insert; lane 1), TLR1_NI (lane 2), TLR1_NS (lane 3), or TLR1_SS (lane 4), or TLR1_SI (lane 5); Western immunoblot probed with an anti-V5 antibody.

Article Snippet: The cells were subsequently incubated with an anti-V5 epitope antibody (Serotec, Oxford, UK) followed by a FITC-conjugated rabbit anti-mouse IgG (Serotec), and thenmounted in mounting medium, containing 25% w/v glycerol, 10% w/v polyvinyl alcohol (MW 30 000–70 000; Sigma-Aldrich), 0.1 M Tris-Cl pH 8.5, and 2.5% 1,4-diazabicyclo-[2.2.2]-octane (DABCO).

Techniques: Immunofluorescence, Expressing, Transfection, Construct, Plasmid Preparation, Control, Western Blot

Journal: bioRxiv

Article Title: Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer

doi: 10.1101/2025.10.06.675352

Figure Lengend Snippet: ( A ) Carboplatin and Triplatin chemical structures. ( B and C ) Cytotoxicity of carboplatin and Triplatin on human OVTOKO and JHOC5 cancer cell lines; 1h treatment. ( D ) Flow cytometry of 200 nM and 50 nM rVAR2-V5 binding to ES2 wt cells, ES2 wt cells + chABC treatment, or Xylt1/Xylt2 KO cells. ( E and F ) Cytotoxicity of carboplatin and Triplatin in ES2 wt and Xylt1/Xylt2 KO cell lines; 1h treatment. ( G and H ). Platinum cellular accumulation in ES2 wt and Xylt1/Xylt2 KO cells treated with 10 µM carboplatin or Triplatin for 1, 2, 4, and 8h. Platinum content was measured by inductively coupled plasma mass spectroscopy (ICP-MS) and normalized by number of cells. ( I and J ). Platinum-DNA adducts in ES2 wt and Xylt1/Xylt2 KO cells treated with 10 µM carboplatin or Triplatin 4, and 8h. ( K and M ) ES2-luc wt and Xylt1/Xylt2 KO tumors were implanted on the left and right flanks mice. Tumors were harvested after 24h treatment with 40 mg/kg i.p Carboplatin or 0.3 mg/kg i.p. Triplatin. Tumors were digested in nitric acid and platinum measured by ICP-MS.

Article Snippet: Bound peptide was detected with mouse anti-V5 antibody (R960-25, ThermoFisher; 1:700 in casein buffer, 45 min, room temperature), followed by Leica post-primary rabbit anti-mouse antibody (8 min) and goat anti-rabbit HRP conjugate (8 min), with PBS washes between each step.

Techniques: Flow Cytometry, Binding Assay, Clinical Proteomics, Mass Spectrometry

Journal: bioRxiv

Article Title: Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer

doi: 10.1101/2025.10.06.675352

Figure Lengend Snippet: Representative images of rVAR2-V5 and H&E OC PDX staining and Qupath analysis. (C) Qupath segmentation of rVAR2-V5 (+), rVAR2-V5 (-), and necrotic tumor area in OC PDX models. (D) Sensitivity of OC PDX models to Triplatin and carboplatin. OC PDX models were treated i.p. with carboplatin (40 mg/kg) or Triplatin (0.3 mg/kg) on days 0, 4 and 8 (orange arrows). * * p<0.01, * * * * p<0.0001, 2-way ANOVA, Tukey

Article Snippet: Bound peptide was detected with mouse anti-V5 antibody (R960-25, ThermoFisher; 1:700 in casein buffer, 45 min, room temperature), followed by Leica post-primary rabbit anti-mouse antibody (8 min) and goat anti-rabbit HRP conjugate (8 min), with PBS washes between each step.

Techniques: Staining

Journal: bioRxiv

Article Title: Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer

doi: 10.1101/2025.10.06.675352

Figure Lengend Snippet: (A) UVA CHTN OC TMA; Representative samples of patient OC subtypes (clinical history unknown) and (B) normal tissues stained with rVAR2-V5 protein. (C) UVA CHTN OC TMA; Percentage of sample area staining positively for rVAR2-V5. Values are representative of the mean of 4 cores per patient sample. (D) UVA CHTN OC TMA; Percentage of TMA samples above the cut-off score. (E) UPenn CCC TMA samples; Percentage of sample area staining positively for rVAR2-V5.

Article Snippet: Bound peptide was detected with mouse anti-V5 antibody (R960-25, ThermoFisher; 1:700 in casein buffer, 45 min, room temperature), followed by Leica post-primary rabbit anti-mouse antibody (8 min) and goat anti-rabbit HRP conjugate (8 min), with PBS washes between each step.

Techniques: Staining

Journal: Journal of Biological Chemistry

Article Title: Amyloid Precursor-like Protein 2 and Sortilin Do Not Regulate the PCSK9 Convertase-mediated Low Density Lipoprotein Receptor Degradation but Interact with Each Other

doi: 10.1074/jbc.m115.647180

Figure Lengend Snippet: FIGURE 1. Exogenous PCSK9 can induce degradation of the LDLR in the absence of APLP2. HepG2 (A) and Huh7 (B) cells were transfected with a control non-target siRNA (Ctrl) or 3 different siRNAs targeting APLP2. Cells wereincubatedovernightwithserum-freeconditionedmedialackingorcon- taining 1 g/ml of PCSK9-V5. HepG2 and Huh7 cell lysates were then sub- jected to Western blotting using LDLR, APLP2, and -actin antibodies. LDLR and APLP2 signals were normalized to that of -actin. C, the input HEK293 conditioned medium was analyzed using mAb-V5 to detect PCSK9-V5. D, duplicate samples of Huh7 cells matching those in panel B were analyzed by FACS to assess the cell surface LDLR levels. Values were normalized to that of the first lane (control non-target siRNA in the absence of PCSK9). Error bars represent S.E. *, p 0.05 (Student’s t test). The data shown here are represen- tative of two to three independent experiments.

Article Snippet: Mouse APLP2 was detected using a rabbit polyclonal antibody kindly provided by Dr. G. Thinakaran (University of Chicago), whereas human APLP2 was detected with either mAb-V5 or a rabbit polyclonal antibody (Novus Biologicals).

Techniques: Transfection, Control, Western Blot

Journal: Journal of Biological Chemistry

Article Title: Amyloid Precursor-like Protein 2 and Sortilin Do Not Regulate the PCSK9 Convertase-mediated Low Density Lipoprotein Receptor Degradation but Interact with Each Other

doi: 10.1074/jbc.m115.647180

Figure Lengend Snippet: FIGURE 4. Sortilin and APLP2 are novel cellular targets of PCSK9. A, overexpressed PCSK9 induces sortilin and APLP2 degradation in HEK293 cells. Triplicate Western blot analyses revealing that both sortilin-Myc and APLP2-V5 expression levels in HEK293 cells were reduced by 90 and 40%, respectively, upon transfection with a PCSK9 plasmid, as compared with a control empty pIRES vector (V). Quantification of sortilin and APLP2 band intensities were normalized against those of -actin. B, HEK293 cells transfected with a cDNA coding for an empty vector control (pIRES; V) or individually with human sortilin or APLP2 tagged at the C terminus with a Myc or V5 epitope, respectively, or together in the absence or presence of a cDNA coding for untagged PCSK9. The following day the cells were washed and then pulsed for 4 h with [35S]Met Cys in the presence or absence of 5 mM NH4Cl. The cells were then extracted and their lysates immunoprecipitated (IP) with a mAb-V5 or mAb-Myc or a polyclonal antibody for PCSK9. The precipitates were separated on an 8% SDS-PAGE. The dried gel was then autoradiographed. Notice the co-precipitation of sortilin and APLP2 in the presence of NH4Cl. These data are representative of at least three independent experiments.

Article Snippet: Mouse APLP2 was detected using a rabbit polyclonal antibody kindly provided by Dr. G. Thinakaran (University of Chicago), whereas human APLP2 was detected with either mAb-V5 or a rabbit polyclonal antibody (Novus Biologicals).

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Control, Immunoprecipitation, SDS Page

Journal: Journal of Biological Chemistry

Article Title: Amyloid Precursor-like Protein 2 and Sortilin Do Not Regulate the PCSK9 Convertase-mediated Low Density Lipoprotein Receptor Degradation but Interact with Each Other

doi: 10.1074/jbc.m115.647180

Figure Lengend Snippet: FIGURE 5. Sortilin, APLP2, and soluble APLP2 are degraded by both PCSK9 and ER-localized PCSK9-KDEL isoforms. HEK293 cells were transfected with indicated DNA amounts of vectors encoding a control protein 7B2, sortilin (no tag), APLP2-V5, soluble APLP2-V5 (sAPLP2-V5), PCSK9-V5, or PCSK9-V5-KDEL, as indicated. After 48 h, lysates and media were analyzed by Western blotting for the indicated proteins. The data show that overexpressed PCSK9 or PCSK9-KDEL induces degradation of sortilin (A), APLP2 (B), and sAPLP2 (C) in HEK293 cells. Quantification of sortilin and APLP2 band intensities were normalized against those of -actin or GAPDH. These data are representative of two independent experiments.

Article Snippet: Mouse APLP2 was detected using a rabbit polyclonal antibody kindly provided by Dr. G. Thinakaran (University of Chicago), whereas human APLP2 was detected with either mAb-V5 or a rabbit polyclonal antibody (Novus Biologicals).

Techniques: Transfection, Control, Western Blot

Journal: Journal of Biological Chemistry

Article Title: Amyloid Precursor-like Protein 2 and Sortilin Do Not Regulate the PCSK9 Convertase-mediated Low Density Lipoprotein Receptor Degradation but Interact with Each Other

doi: 10.1074/jbc.m115.647180

Figure Lengend Snippet: FIGURE 6. Co-expression of sortilin, APLP2, or both with PCSK9 has no major effect on LDLR degradation. Huh7 cells were transfected with a total of 3 g using 1 g of each vector encoding for either a control protein 7B2 (), sortilin (), APLP2 (), or PCSK9 (), as indicated. After 48 h, lysates were analyzed by Western blotting for expression of the LDLR, sortilin-Myc, APLP2- V5, intracellular pro- and mature-PCSK9-V5, and -actin. Media were ana- lyzed for secreted endogenous and overexpressed PCSK9-V5 using a rabbit polyclonal human PCSK9 antibody. Quantification of LDLR expression was normalized against that of -actin. These data are representative of at least 3 different experiments showing similar results.

Article Snippet: Mouse APLP2 was detected using a rabbit polyclonal antibody kindly provided by Dr. G. Thinakaran (University of Chicago), whereas human APLP2 was detected with either mAb-V5 or a rabbit polyclonal antibody (Novus Biologicals).

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Western Blot

Journal: Journal of Biological Chemistry

Article Title: Amyloid Precursor-like Protein 2 and Sortilin Do Not Regulate the PCSK9 Convertase-mediated Low Density Lipoprotein Receptor Degradation but Interact with Each Other

doi: 10.1074/jbc.m115.647180

Figure Lengend Snippet: FIGURE 7. Sortilin binds APLP2. A, schematic diagram of sortilin and APLP2 fused to the G. princeps luciferase half-domains, Gluc-1 and Gluc-2, respectively. B, heat map generated by G. princeps luciferase complementation assay showing the interaction profile of 36 protein pairs. Normalized luminescence ratio ranging from strong to null interactions is displayed on a light blue to black scale. C, validation of sortilin-Myc and APLP2-V5 interaction was confirmed by co-expression in HEK293 cells and immunoprecipitation with mAb-Myc followed by Western blotting (WB) using a mAb-Myc or mAb-V5.

Article Snippet: Mouse APLP2 was detected using a rabbit polyclonal antibody kindly provided by Dr. G. Thinakaran (University of Chicago), whereas human APLP2 was detected with either mAb-V5 or a rabbit polyclonal antibody (Novus Biologicals).

Techniques: Luciferase, Generated, Biomarker Discovery, Expressing, Immunoprecipitation, Western Blot