Journal: Cell reports

Article Title: The Borrelia burgdorferi VlsE Lipoprotein Prevents Antibody Binding to an Arthritis-Related Surface Antigen

doi: 10.1016/j.celrep.2020.02.081

Figure Lengend Snippet:

Article Snippet: Anti-VlsE IgG , Rockland , Cat# 200-401-C33; RRID:AB_10924424.

Techniques: Recombinant, Blocking Assay, Western Blot, Staining, Purification, DNA Purification, Software, Luminex, Imaging, Flow Cytometry

Journal: Cell reports

Article Title: The Borrelia burgdorferi VlsE Lipoprotein Prevents Antibody Binding to an Arthritis-Related Surface Antigen

doi: 10.1016/j.celrep.2020.02.081

Figure Lengend Snippet:

Article Snippet: VlsE Control Protein , Rockland , Cat# 000-001-C33.

Techniques: Virus, Recombinant, Blocking Assay, Western Blot, Staining, Control, Purification, DNA Purification, Software, Luminex, Imaging, Flow Cytometry, Membrane

Journal: The Journal of Neuroscience

Article Title: Dynamic Partitioning of Synaptic Vesicle Pools by the SNARE-Binding Protein Tomosyn

doi: 10.1523/jneurosci.1297-16.2016

Figure Lengend Snippet: Figure3. Tomo1servesascatalyticsubstrateforneuralactivity-sensitiveCdk5.A1,RepresentativeimmunofluorescentimagesofSynapsin1(cyan,top)andphosphorylatedCdk5(pCdk5;heat scale,bottom)instraightenedaxonsegmentsfromneuronstreated(30min)withCSA(50M),Rosco(100M),orcontrol(DMSO,0.5%).A2,AveragedpCdk5immunofluorescenceintensityrelative to vehicle control. A3, Cumulative frequency of pCdk5 intensity across boutons shows that inhibition of Cdk5 by Rosco significantly reduces pCdk5 immunoreactivity, whereas CSA treatment enhances pCdk5 relative to control. B1–B3, pCdk5 immunofluorescence images comparing chronic silencing of neural activity by TTX (24 h, 1 M) with control. Averaged pCdk5 intensity (B2) and cumulativefrequencyofpCdk5(B3)intensitydemonstratethatchronicdampeningofneuralactivitysignificantlyreducesCdk5activation.A,B,Analysiswasperformedon45images/preparation per condition and repeated on 3 neuronal preparations. C, Immunoblots showing coprecipitation of Tomo1 and Cdk5 from cultured hippocampal neuron lysates regardless of primary target of IP (Tomo1 or Cdk5). D, In vitro phosphorylation of affinity-purified Tomo1 by Cdk5/p25 as measured by 32P labeling. 32P-radioactive signal (top) and anti-Tomo1 immunoreactivity (bottom) on Western blot corresponding to the following conditions: absence of Cdk5/p25 kinase (H2O), Cdk5/p25 kinase (Cdk5), and Cdk5/p25 1 mM Rosco (Cdk5 Rosco). *p 0.05.

Article Snippet: For in vitro Cdk5/p25 phosphorylation reactions, 11.5 g of mTomo1 on streptavidin beads was rinsed in phosphorylation buffer containing the following (in mM): 25 MOPS, pH 7.2, 12.5 -glycerol phosphate, 25 MgCl2, 5 EGTA, 2 EDTA, 0.25 DTT, and 250 g of active Cdk5/p25 kinase (Signal Chem, C33–10G).

Techniques: Control, Inhibition, Immunofluorescence, Activity Assay, Western Blot, Cell Culture, In Vitro, Phospho-proteomics, Affinity Purification, Labeling

Journal: bioRxiv

Article Title: t-Darpp is an elongated monomer that binds to calcium and is phosphorylated by cyclin-dependent kinases 1 and 5

doi: 10.1101/152272

Figure Lengend Snippet: CDK1 and CDK5 phosphorylate t-Darpp in vitro . In vitro kinase assay performed using recombinant t-Darpp (tDp) and CDK1 (A) or CDK5 (B). Reactions were performed over a period of 1 h with aliquots taken at 0, 5, 15, 30 and 60 min. Inhibitors, RO-3306 and roscovitine were used as controls to inhibit CDK1 and CDK5, respectively. The T39-phosphospecific antibody was used to show phosphorylation at T39 of t-Darpp. H62 was used as a loading control antibody to show the relative level of recombinant t-Darpp in each lane.

Article Snippet: CDK1 (C22-18G) and CDK5 (C33-10G-05) were purchased from Signal Chem Pharmaceuticals (Richmond BC, Canada).

Techniques: In Vitro, Kinase Assay, Recombinant, Phospho-proteomics, Control

Journal: Frontiers in Immunology

Article Title: CD82-associated exhausted CD8 + T cells define prognosis and immunotherapy resistance in colon cancer

doi: 10.3389/fimmu.2025.1731154

Figure Lengend Snippet: Expression distribution of CD82 in CD8 + T cell subsets analyzed by scRNA-seq. (A) UMAP visualization of cell types based on scRNA-seq data obtained from the GEO database. (B) Dot plot illustrating CD82 expression levels across different cell types in tumor and normal tissues from patients with colon cancer. (C) UMAP visualization of CD8 + T cell subsets based on scRNA-seq data obtained from the GEO database. (D) Dot plot illustrating CD82 expression levels across distinct CD8 + T cell subsets in tumor and normal tissues from patients with colon cancer.

Article Snippet: The following antibodies are used: CD82 (clone 1E7A4, 1:1000 dilution, Catalog No.: 66803-1-lg, Proteintech, USA), CD8 (clone 1G2B10, 1:2000 dilution, Cat. No.: 66868-1-Ig, Proteintech, USA), TIM-3 (clone D5D5RTM, 1:100 dilution, Catalog No.: 45208S, CST, USA), TCF1 (clone C63D9, 1:200 dilution, Catalog No.: 2203S, CST, USA), PD-1 (clone UMAB199, 1:100 dilution, Cat. No.: ZM0381, ZSGB-BIO, China), and PANCK (clone PDH09-10, 1:2000 dilution, Cat. No.: HA601138 , HUABio, China).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: CD82-associated exhausted CD8 + T cells define prognosis and immunotherapy resistance in colon cancer

doi: 10.3389/fimmu.2025.1731154

Figure Lengend Snippet: Correlation analysis of CD82 expression with T cell exhaustion markers in TCGA-COAD. (A–H) Scatter plots illustrating the correlations between CD82 expression and T cell exhaustion–associated molecules in colon cancer tissues: (A) CTLA4, (B) FOXP3, (C) TIGIT, (D) CD8A, (E) MRC1 (CD206), (F) PDCD1 (PD-1), (G) HAVCR2 (TIM-3), and (H) LAG3. Correlations are assessed using Pearson’s correlation analysis (all P < 0.001).

Article Snippet: The following antibodies are used: CD82 (clone 1E7A4, 1:1000 dilution, Catalog No.: 66803-1-lg, Proteintech, USA), CD8 (clone 1G2B10, 1:2000 dilution, Cat. No.: 66868-1-Ig, Proteintech, USA), TIM-3 (clone D5D5RTM, 1:100 dilution, Catalog No.: 45208S, CST, USA), TCF1 (clone C63D9, 1:200 dilution, Catalog No.: 2203S, CST, USA), PD-1 (clone UMAB199, 1:100 dilution, Cat. No.: ZM0381, ZSGB-BIO, China), and PANCK (clone PDH09-10, 1:2000 dilution, Cat. No.: HA601138 , HUABio, China).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: CD82-associated exhausted CD8 + T cells define prognosis and immunotherapy resistance in colon cancer

doi: 10.3389/fimmu.2025.1731154

Figure Lengend Snippet: Expression of CD82 in colon cancer tissue microarrays (TMA) and its prognostic significance. (A) Representative multiplex immunohistochemical (mIHC) staining and single-channel images of colon cancer TMAs (DAPI, blue; CK, purple; CD8, cyan; CD82, white). (B–D) Comparisons between normal and tumor tissues: (B) infiltration density of CD8 + T cells, (C) infiltration density of D82 + T cells, and (D) proportion of the CD8 + CD82 + subset within total CD8 + T cells. (E–G) Comparisons between epithelial and stromal regions within tumor tissues: (E) infiltration density of CD8 + T cells, (F) infiltration density of CD82 + T cells, and (G) proportion of the CD8 + CD82 + subset within total CD8 + T cells. (H–J) Kaplan–Meier survival curves for colon cancer patients stratified by: (H) CD8 + T cell infiltration density, (I) CD82 + T cell infiltration density, and (J) proportion of the CD8 + CD82 + subset within total CD8 + T cells. Optimal cutoff values for high and low expression groups are determined using X-tile software. P values for survival differences are calculated using the Kaplan–Meier method, and HR with 95%CI are estimated by univariate Cox proportional hazards regression analysis. ***P < 0.001; ****P < 0.0001.

Article Snippet: The following antibodies are used: CD82 (clone 1E7A4, 1:1000 dilution, Catalog No.: 66803-1-lg, Proteintech, USA), CD8 (clone 1G2B10, 1:2000 dilution, Cat. No.: 66868-1-Ig, Proteintech, USA), TIM-3 (clone D5D5RTM, 1:100 dilution, Catalog No.: 45208S, CST, USA), TCF1 (clone C63D9, 1:200 dilution, Catalog No.: 2203S, CST, USA), PD-1 (clone UMAB199, 1:100 dilution, Cat. No.: ZM0381, ZSGB-BIO, China), and PANCK (clone PDH09-10, 1:2000 dilution, Cat. No.: HA601138 , HUABio, China).

Techniques: Expressing, Multiplex Assay, Immunohistochemical staining, Staining, Software

Journal: Frontiers in Immunology

Article Title: CD82-associated exhausted CD8 + T cells define prognosis and immunotherapy resistance in colon cancer

doi: 10.3389/fimmu.2025.1731154

Figure Lengend Snippet: Co-expression of CD82 and exhaustion markers in colon cancer TMA and their prognostic significance. (A) Representative mIHC staining and single-channel images (DAPI, blue; CK, purple; CD8, cyan; CD82, white; TIM-3, green; PD-1, orange; TCF1, red). (B–D) Comparisons between normal and tumor tissues: (B) proportion of CD8 + CD82 + PD-1 + cells within CD8 + PD-1 + T cells, (C) proportion of CD8 + CD82 + PD-1 + TCF1 + cells within CD8 + PD-1 + TCF1 + T cells, and (D) proportion of CD8 + CD82 + TIM-3 + PD-1 + cells within CD8 + TIM-3 + PD-1 + T cells. (E–G ) Comparisons between epithelial and stromal regions within tumor tissues: (E) proportion of CD8 + CD82 + PD-1 + cells within CD8 + PD-1 + T cells, (F) proportion of CD8 + CD82 + PD-1 + TCF1 + cells within CD8 + PD-1 + TCF1 + T cells, and (G) proportion of CD8 + CD82 + TIM-3 + PD-1 + cells within CD8 + TIM-3 + PD-1 + T cells. (H–J) Kaplan–Meier survival curves for colon cancer patients stratified by: (H) proportion of CD8 + CD82 + PD-1 + cells within CD8 + PD-1 + T cells, (I) proportion of CD8 + CD82 + PD-1 + TCF1 + cells within CD8 + PD-1 + TCF1 + T cells, and (J) proportion of CD8 + CD82 + TIM-3 + PD-1 + cells within CD8 + TIM-3 + PD-1 + T cells. Optimal cutoff values for high and low expression groups are determined using X-tile software. P values for survival differences are calculated using the Kaplan–Meier method, and HR with 95%CI are estimated by univariate Cox proportional hazards regression analysis. *P < 0.05; ****P < 0.0001.

Article Snippet: The following antibodies are used: CD82 (clone 1E7A4, 1:1000 dilution, Catalog No.: 66803-1-lg, Proteintech, USA), CD8 (clone 1G2B10, 1:2000 dilution, Cat. No.: 66868-1-Ig, Proteintech, USA), TIM-3 (clone D5D5RTM, 1:100 dilution, Catalog No.: 45208S, CST, USA), TCF1 (clone C63D9, 1:200 dilution, Catalog No.: 2203S, CST, USA), PD-1 (clone UMAB199, 1:100 dilution, Cat. No.: ZM0381, ZSGB-BIO, China), and PANCK (clone PDH09-10, 1:2000 dilution, Cat. No.: HA601138 , HUABio, China).

Techniques: Expressing, Staining, Software

Journal: Frontiers in Immunology

Article Title: CD82-associated exhausted CD8 + T cells define prognosis and immunotherapy resistance in colon cancer

doi: 10.3389/fimmu.2025.1731154

Figure Lengend Snippet: scRNA-seq analysis and expression distribution of CD82 in CD8 + T-cell subpopulations following immunotherapy. (A, B) UMAP visualization of cell types (A) and CD82 expression distribution (B) based on scRNA-seq data from GEO datasets of colon cancer patients treated with immunotherapy. (C) Dot plot showing differences in CD82 expression levels across cell types between pathological complete response (pCR) and non-pCR groups after immunotherapy. (D, E) UMAP visualization of CD8 + T-cell subsets (D) and corresponding CD82 expression distribution (E) . (F) Scatter plot comparing CD82 expression levels among distinct CD8 + T-cell subsets between pCR and non-pCR groups post-immunotherapy. (G) Violin plot illustrating CD82 expression differences among CD8 + T-cell subsets between pCR and non-pCR groups following immunotherapy.

Article Snippet: The following antibodies are used: CD82 (clone 1E7A4, 1:1000 dilution, Catalog No.: 66803-1-lg, Proteintech, USA), CD8 (clone 1G2B10, 1:2000 dilution, Cat. No.: 66868-1-Ig, Proteintech, USA), TIM-3 (clone D5D5RTM, 1:100 dilution, Catalog No.: 45208S, CST, USA), TCF1 (clone C63D9, 1:200 dilution, Catalog No.: 2203S, CST, USA), PD-1 (clone UMAB199, 1:100 dilution, Cat. No.: ZM0381, ZSGB-BIO, China), and PANCK (clone PDH09-10, 1:2000 dilution, Cat. No.: HA601138 , HUABio, China).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: CD82-associated exhausted CD8 + T cells define prognosis and immunotherapy resistance in colon cancer

doi: 10.3389/fimmu.2025.1731154

Figure Lengend Snippet: Single-cell regulatory network inference and clustering analysis of CD8 + T cells. (A) Heatmap showing regulon activity (AUCell AUC values) across CD8 + T-cell subsets. (B) Differentially expressed transcriptional regulators between pCR and non-pCR groups predicted to bind the promoter or enhancer regions of the CD82 gene. (C) Comparison of mRNA expression levels of candidate regulatory factors between pCR and non-pCR groups.

Article Snippet: The following antibodies are used: CD82 (clone 1E7A4, 1:1000 dilution, Catalog No.: 66803-1-lg, Proteintech, USA), CD8 (clone 1G2B10, 1:2000 dilution, Cat. No.: 66868-1-Ig, Proteintech, USA), TIM-3 (clone D5D5RTM, 1:100 dilution, Catalog No.: 45208S, CST, USA), TCF1 (clone C63D9, 1:200 dilution, Catalog No.: 2203S, CST, USA), PD-1 (clone UMAB199, 1:100 dilution, Cat. No.: ZM0381, ZSGB-BIO, China), and PANCK (clone PDH09-10, 1:2000 dilution, Cat. No.: HA601138 , HUABio, China).

Techniques: Activity Assay, Comparison, Expressing

Journal: bioRxiv

Article Title: Melanoblast transcriptome analysis reveals novel pathways promoting melanoma metastasis

doi: 10.1101/721712

Figure Lengend Snippet: a , Screen of known melanoma metastasis suppressor expression following KDELR3 knockdown (3 days post knockdown). P, Parental; C, siControl; K3, si KDELR3. b , qPCR of KAI1 RNA expression ( CD82 gene) in siRNA knockdown cells (indicated), 3 days post knockdown. c-e , KAI1 protein ( c ) and RNA ( d - e ) expression in 1205Lu cells transfected with CD82/KAI1 overexpression (KAI1) or PCMV6-AC control vector (Vec.), KDELR3 transcript 1with DDK tag (K3_1), KDELR3 transcript 2 with DDK tag (K3_2), or PCMV6 control vector (Vec.1). Harvested 3 days post transfection. Equal protein amounts subjected to immunoblot analysis with an anti-KAI1 and anti-DDK antibody and anti-VINCULIN loading control ( c ). f , 1205Lu cells parental (P), and 1205Lu cells transiently transfected with control siRNA (C), and KDELR3 siRNA (K3), harvested 3 days post transfection and equal protein amounts subjected to immunoblot analysis with an anti-KAI1 and anti-gp78 antibody. g , KAI1 protein expression in siRNA knockdown (indicated) 1205Lu cells harvested 3 days post transfection and treated with de-glycosylation enzymes (De-G). h , qPCR of gp78 RNA expression ( AMFR gene) in siRNA knockdown cells (indicated), 3 days post knockdown. f , g , Anti-vinculin antibody used to control for protein loading. i , qPCR of KDELR3 RNA expression in siRNA knockdown cells (indicated), 4 days post knockdown j , Co-immunoprecipitation of endogenous gp78 and mCherry tagged gp78 (gp78-mCh) with FLAG-tagged KDELR3 (K3-DDK) in stably transduced 1205Lu cells. Red line, gp78 expression. k , pol2 > KDELR3 -GFP (green) co-localizes with pol2> gp78 -mCherry (red) in 1205Lu metastatic melanoma cells. Scale bars, 50 µm. l , Schematic of the KDELR3-KAI1 axis in melanoma metastasis. a - f , h , j - k , Representative of three independent experiments. g , i , Representative of two independent experiments.

Article Snippet: For exogenous over-expression of CD82 and KDELR3 genes the following expression plasmids were used: CD82 transcript variant 1 (NM_002231) Human Untagged Clone (Origene, CAT#: SC324395), pCMV6-AC Tagged Cloning mammalian vector with non-tagged expression (Origene, CAT#: PS100020), KDELR3 transcript variant 2 (NM_016657) Human Myc-DDK-tagged ORF Clone (Origene, CAT#: RC216726), KDELR3 transcript variant 1 (NM_006855) Human Myc-DDK-tagged ORF Clone (Origene, CAT#: RC201571), pCMV6-Entry Tagged Cloning mammalian vector with C-terminal Myc-DDK Tag (Origene, CAT#: PS100001).

Techniques: Expressing, RNA Expression, Transfection, Over Expression, Plasmid Preparation, Western Blot, Immunoprecipitation, Stable Transfection