jurkat t cells  (ATCC)

Journal: Oncology Letters

Article Title: BUD23 is associated with malignancy and correlates with immune infiltration in NSCLC

doi: 10.3892/ol.2026.15608

Figure Lengend Snippet: BUD23 knockdown suppresses the proliferative and migration of NSCLC cells. (A) RT-qPCR was used to quantify BUD23 mRNA levels in HBE cells and a panel of NSCLC cell lines. Validation of BUD23 knockdown efficiency in (B) A549 and (C) H1299 cells by RT-qPCR. Assessment of cell viability in (D) A549 and (E) H1299 cells via CCK-8 assay and cell migration in (F) A549 and (G) H1299 cells by wound healing assay (magnification, ×10). CCK-8 assays of (H) A549 and (I) H1299 cells 24 h after BUD23 knockdown with or without subsequent co-culture with Jurkat T cells for 48 h in Transwell chambers. (J) Quantification of apoptosis in A549 and H1299 cells via Annexin V/PI flow cytometry. **P<0.01 and ***P<0.001 vs. HBE, NC, Con or as indicated. NSCLC, non-small cell lung cancer; RT-qPCR, reverse transcription-quantitative PCR; HBE, human bronchial epithelial; CCK-8, Cell Counting Kit-8; NC, negative control; Con, control; Si1/2, small interfering RNA targeting BUD23.

Article Snippet: The HBE [full name: HBE4-E6/E7 (Human Bronchial Epithelial Cells; cat. no. CRL-2078)] cell line, A549 lung adenocarcinoma cell line (cat. no. CCL-185), H1299 lung large cell carcinoma cell line (cat. no. CRL-5803), H460 lung large cell carcinoma cell line (cat. no. HTB-177) and Jurkat T cells (cat. no. TIB-152; a childhood T acute lymphoblastic leukemia T-cell line), were purchased from the American Type Culture Collection.

Techniques: Knockdown, Migration, Quantitative RT-PCR, Biomarker Discovery, CCK-8 Assay, Wound Healing Assay, Co-Culture Assay, Flow Cytometry, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting, Negative Control, Control, Small Interfering RNA

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: In activated Jurkat cells, human Galectin-10 produced marked Annexin V and Annexin V/PI positivity at 24 h.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced

Journal: Current Protocols

Article Title: Engineering, Expression, Purification, and Application of Glycosaminoglycan‐Specific Antibodies

doi: 10.1002/cpz1.70358

Figure Lengend Snippet: Use of anti‐HS scFvs as staining reagents for flow cytometry. ( A‐C ) Workflow, gating strategy, and representative histograms for FACS analyses of ( A ) live Jurkat E6‐1 cells stained with anti‐HS scFv‐H HS001 and PE‐conjugated anti‐His antibody (data collected on Sony SA3800 spectral analyzer); ( B ) Vero cells stained with anti‐HS scFv‐H HS001, then fixed and detected using AlexaFluor647‐conjugated anti‐human IgG antibody (data collected on Novocyte Quanteon in AlexaFluor647 channel); and ( C ) live Vero cells stained with TAMRA‐conjugated anti‐HS scFvs (data collected on Novocyte Quanteon in the PE/TAMRA channel). HS034 is a negative control scFv and has no known epitope. scFv staining observed in live, single‐cell populations.

Article Snippet: Figure shows representative data as heatmaps for % live cells showing scFv staining obtained using the FACS staining protocol detailed in Basic Protocol for two cell lines: HEK293 cells (ATCC, CRL‐1573, maintained in DMEM supplemented with l ‐glutamine, glucose, sodium pyruvate, and 10% FBS) and Jurkat E6‐1 cells (ATCC, TIB‐152, maintained in IMDM with GlutaMAX and 10% FBS).

Techniques: Staining, Flow Cytometry, Negative Control, Single Cell

Journal: Current Protocols

Article Title: Engineering, Expression, Purification, and Application of Glycosaminoglycan‐Specific Antibodies

doi: 10.1002/cpz1.70358

Figure Lengend Snippet: Data for qualitative validation of the anti‐HS scFv‐H panel. Representative data for % live cells showing scFv staining observed for HEK293 and Jurkat E6‐1 cells, obtained following indirect FACS staining as in Basic Protocol ).

Article Snippet: Figure shows representative data as heatmaps for % live cells showing scFv staining obtained using the FACS staining protocol detailed in Basic Protocol for two cell lines: HEK293 cells (ATCC, CRL‐1573, maintained in DMEM supplemented with l ‐glutamine, glucose, sodium pyruvate, and 10% FBS) and Jurkat E6‐1 cells (ATCC, TIB‐152, maintained in IMDM with GlutaMAX and 10% FBS).

Techniques: Biomarker Discovery, Staining

Journal: bioRxiv

Article Title: Exosome-like biogenesis from the Golgi releases extracellular vesicles lacking conventional tetraspanins that mediate immune evasion in cancer

doi: 10.64898/2026.05.13.719612

Figure Lengend Snippet: a. Jurkat T cell proliferation following treatment with EVs derived from NSCLC cell lines (A549, HCC827, and H358) at 5, 10, and 20 µg for 96 hr. Proliferation was normalized to PBS-treated Jurkat T cells (dashed line) (mean ± SD, two-way ANOVA with Dunnett’s multiple comparisons test; n=4). b. Left: Representative flow cytometry dot plot showing % cells positive for propidium iodide staining (necrosis marker) in Jurkat T cells treated with 25 µg of H358 EVs or PBS. Right: Representative histogram of CD69 expression (activation marker) under the same conditions. Mean values indicated above the histogram. c. Top: Representative histograms of eFluor™ 450 dye dilution in primary human CD3+, CD3+CD4+, and CD3+CD8+ T cells following treatment with H358 EVs for 96 hr at indicated doses. Reduced dye intensity indicated increased proliferation. Unactivated T cells and volume-matched PBS-treated T cells served as negative and positive controls, respectively. Full gating strategy shown in Fig. S6a. Bottom: Quantification of proliferating T cells (cells left of dashed line) expressed as a percentage relative to PBS control (mean ± SD, one-way ANOVA with Dunnett’s multiple comparisons test; n=6). Primary T cells from a single donor treated with six different H358 EV preps. d. Representative images of human bronchial epithelial cells treated with A549 or H358 EVs (7.5 and 15 µg; 96 hr) compared with PBS control. Bottom: Quantification of cell number normalized to PBS-treated control (mean ± SD, one-way ANOVA with Dunnett’s multiple comparisons test; n=3).

Article Snippet: A549 (CCL-185TM, RRID:CVCL_0023), HCC827 (CRL-2868TM, RRID:CVCL_2063) and H358 (CRL-5807TM, RRID:CVCL_1559) were purchased from ATCC, and Jurkat T cells (RRID:CVCL_0065) were provided by Dr. Kazemian (Purdue University).

Techniques: Derivative Assay, Flow Cytometry, Staining, Marker, Expressing, Activation Assay, Control

Journal: bioRxiv

Article Title: Exosome-like biogenesis from the Golgi releases extracellular vesicles lacking conventional tetraspanins that mediate immune evasion in cancer

doi: 10.64898/2026.05.13.719612

Figure Lengend Snippet: a. Representative immunoblot showing canonical EV markers CD81 and CD63 in H358 EV lysate (5 µg total protein) compared to H358 whole-cell lysate (5 and 25 µg). GAPDH confirms minimal cellular contamination of isolated EVs. b. Immunoblot of pull-down (PD) and flow-through (FT) fractions following CD81- and CD63-immunoprecipitation of H358 EVs. IgG immunoprecipitation served as a control. c. Jurkat T cell proliferation following treatment with total H358 EVs or EVs depleted of CD81+ or CD63+ vesicles (96 hr). EV input was normalized to 5 µg total protein. Proliferation was quantified by eFluor™ 450 dye dilution (mean ± SD; unpaired t-test; n=3). d. Schematic illustrating Brefeldin A (BFA) treatment to inhibit Golgi trafficking and assess its effect on EV secretion. e. Quantification of EV secretion from A549 and H358 cells following 48 hr treatment with BFA (8 ng/mL). EV number is shown per 10 cells. Untreated cells served as a control (mean ± SD; unpaired t-test; n=3). f. Jurkat T cell proliferation following treatment with equal volumes of EVs derived from untreated or BFA-treated H358 cells. Proliferation was normalized to PBS-treated Jurkat T cells (mean ± SD; unpaired t-test; n=4). g. Immunoblot showing expression of the trans-Golgi marker TGOLN2 and canonical EV markers in EV lysates (10 µg total protein) from HCC827, A549, and H358 cells. h. Immunoblot of PD and FT fractions following TGOLN2 or CD81 immunoprecipitation of H358 EVs. IgG immunoprecipitation served as a control. i. Top: Representative histogram of primary human T cell proliferation following treatment with total H358 EVs or EVs depleted of CD81+ or CD63+ vesicles (96 hr), assessed by eFluor™ 450 dye dilution. Bottom: Quantification of proliferating T cells (cells left of dashed line) relative to PBS-treated controls (mean ± SD; one-way ANOVA with Dunnett’s post hoc, n=6). j. Immunoblot of TGOLN2 in EV and whole-cell lysates from TGOLN2-overexpressing A549 cells (A549-TGOLN2) and parent A549 cells. CD81 and GAPDH confirm EV enrichment and purity. k. Jurkat T cell proliferation following treatment with equal numbers of EVs from parental A549 or A549-TGOLN2 cells. Proliferation normalized to PBS-treated controls (mean ± SD; unpaired t-test; n=4). l. Immunoblot of PD and FT fractions following TGOLN2-immunoprecipitation of EVs derived from A549-TGOLN2 cells. CD9 is shown as a canonical EV marker. IgG served as a control. m. Jurkat T cell proliferation following treatment with total A549-TGOLN2 EVs, mock-depleted EVs (IgG), or TGOLN2-depleted EVs (96 hr; 10 and 25 µL input volumes). Proliferation normalized to PBS-treated Jurkat T cells (mean ± SD; unpaired t-test; n=4). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

Article Snippet: A549 (CCL-185TM, RRID:CVCL_0023), HCC827 (CRL-2868TM, RRID:CVCL_2063) and H358 (CRL-5807TM, RRID:CVCL_1559) were purchased from ATCC, and Jurkat T cells (RRID:CVCL_0065) were provided by Dr. Kazemian (Purdue University).

Techniques: Western Blot, Isolation, Immunoprecipitation, Control, Derivative Assay, Expressing, Marker