Journal: The Journal of biological chemistry

Article Title: Anti-IgM-mediated regulation of c-myc and its possible relationship to apoptosis.

doi: 10.1074/jbc.271.31.18875

Figure Lengend Snippet: FIG. 8. Cell growth in various Burkitt’s lymphoma cell lines treated with anti-IgM. Daudi and Ramos cells were seeded at a density of 1 3 105 cells/ml in 200 ml in a 96-well microtiter plate and treated (10 mg/ml anti-IgM). Cell numbers were determined at indi- cated times using a Thomas hemacytometer. (E) untreated cells; (G) anti-IgM-treated. Raji, CA46, Namalwa, and ST486 cells were seeded at a density of 5 3 104 cells/ml in 200 ml in 96-well microtiter plates and treated (10 mg/ml anti-IgM). Cell numbers at the indicated times were determined by an ELISA method. (E) untreated cells; (G) anti- IgM-treated.

Article Snippet: Daudi (ATCC CCL 213), Ramos (ATCC CRL 1596), Raji (ATCC CCL 86), Namalwa (ATCC CRL 1432), ST486 (ATCC CRL 1647), CA46 (ATCC CRL 1648) cell lines and GAPDH plasmid (ATCC 57090) were obtained from American Type Culture Collection (Rockville, MD).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: The Journal of biological chemistry

Article Title: Anti-IgM-mediated regulation of c-myc and its possible relationship to apoptosis.

doi: 10.1074/jbc.271.31.18875

Figure Lengend Snippet: FIG. 11. The role of c-myc in regulating cell growth, and the effect of EBV proteins in preventing apoptosis. Burkitt’s lym- phoma cells which are p53 mutant (103) and express high level of c-myc (10, 17, 105) can grow rapidly in vitro. Cross-linking of B cell antigen receptor by anti-IgM in these cells induces down-regulation of c-myc mRNA as well as Myc protein. Reduction of the c-myc level in these cells will propel the cells to exit the cell cycle. In the absence of EBV proteins (EBNAs, LMPs, and BHRF-1 (bcl-2 homologue)), the cells will result in apoptosis (e.g. Ramos and ST486 cells). However, in the presence of EBV proteins, apoptosis is blocked resulting in growth arrest (e.g. Daudi, Raji, and Namalwa cells). Addition of antisense c-myc oligonu- cleotides to these cells demonstrated a similar result to that seen with anti-IgM.

Article Snippet: Daudi (ATCC CCL 213), Ramos (ATCC CRL 1596), Raji (ATCC CCL 86), Namalwa (ATCC CRL 1432), ST486 (ATCC CRL 1647), CA46 (ATCC CRL 1648) cell lines and GAPDH plasmid (ATCC 57090) were obtained from American Type Culture Collection (Rockville, MD).

Techniques: Mutagenesis, In Vitro

Journal: The Journal of biological chemistry

Article Title: Anti-IgM-mediated regulation of c-myc and its possible relationship to apoptosis.

doi: 10.1074/jbc.271.31.18875

Figure Lengend Snippet: FIG. 8. Cell growth in various Burkitt’s lymphoma cell lines treated with anti-IgM. Daudi and Ramos cells were seeded at a density of 1 3 105 cells/ml in 200 ml in a 96-well microtiter plate and treated (10 mg/ml anti-IgM). Cell numbers were determined at indi- cated times using a Thomas hemacytometer. (E) untreated cells; (G) anti-IgM-treated. Raji, CA46, Namalwa, and ST486 cells were seeded at a density of 5 3 104 cells/ml in 200 ml in 96-well microtiter plates and treated (10 mg/ml anti-IgM). Cell numbers at the indicated times were determined by an ELISA method. (E) untreated cells; (G) anti- IgM-treated.

Article Snippet: Daudi (ATCC CCL 213), Ramos (ATCC CRL 1596), Raji (ATCC CCL 86), Namalwa (ATCC CRL 1432), ST486 (ATCC CRL 1647), CA46 (ATCC CRL 1648) cell lines and GAPDH plasmid (ATCC 57090) were obtained from American Type Culture Collection (Rockville, MD).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: The Journal of biological chemistry

Article Title: Anti-IgM-mediated regulation of c-myc and its possible relationship to apoptosis.

doi: 10.1074/jbc.271.31.18875

Figure Lengend Snippet: FIG. 11. The role of c-myc in regulating cell growth, and the effect of EBV proteins in preventing apoptosis. Burkitt’s lym- phoma cells which are p53 mutant (103) and express high level of c-myc (10, 17, 105) can grow rapidly in vitro. Cross-linking of B cell antigen receptor by anti-IgM in these cells induces down-regulation of c-myc mRNA as well as Myc protein. Reduction of the c-myc level in these cells will propel the cells to exit the cell cycle. In the absence of EBV proteins (EBNAs, LMPs, and BHRF-1 (bcl-2 homologue)), the cells will result in apoptosis (e.g. Ramos and ST486 cells). However, in the presence of EBV proteins, apoptosis is blocked resulting in growth arrest (e.g. Daudi, Raji, and Namalwa cells). Addition of antisense c-myc oligonu- cleotides to these cells demonstrated a similar result to that seen with anti-IgM.

Article Snippet: Daudi (ATCC CCL 213), Ramos (ATCC CRL 1596), Raji (ATCC CCL 86), Namalwa (ATCC CRL 1432), ST486 (ATCC CRL 1647), CA46 (ATCC CRL 1648) cell lines and GAPDH plasmid (ATCC 57090) were obtained from American Type Culture Collection (Rockville, MD).

Techniques: Mutagenesis, In Vitro

Journal: The Journal of infectious diseases

Article Title: Aberrant Humoral Immune Responses in Neurosyphilis: CXCL13/CXCR5 Play a Pivotal Role for B-Cell Recruitment to the Cerebrospinal Fluid.

doi: 10.1093/infdis/jix233

Figure Lengend Snippet: Figure 1. Enrichment of CD19+ B cells and memory B cells in the CSF of NS patients. A, Gating strategies. B, Representative flow cytometry analysis of the frequency of CSF total CD19+ B cells and subsets of CD19+ B cells between non-NS and NS patients (first row). Bar charts of the frequency of B-cell subsets of patients with asymptomatic and symptomatic NS (second row). Comparison of the proportion of CSF total CXCR5+ B cells in different groups (third row). C, Serial changes of CSF total CD19+ B cells in the patient with NS following first and second treatment. D, Correlation of frequency of CD19+ B cells and CSF protein levels, CSF WBC counts, or CSF VDRL titer. Abbreviations: CSF, cerebrospinal fluid; NS, neurosyphilis; VDRL, Venereal Disease Research Laboratory; WBC, white blood cell.

Article Snippet: Paraffin-embedded brain biopsy (4 μm) was stained with appropriate concentration of primary antibodies against CXCL13, CXCR5, CD20, CD3, CD35, CD138 (Maixin-Bio, Fujian, China), and spirochete (Biocare Medical, Pacheco, CA), then incubated with streptavidin-biotin complex system according to manufacturer’s instructions (Boster, Pleasanton, CA).

Techniques: Flow Cytometry, Comparison

Journal: The Journal of infectious diseases

Article Title: Aberrant Humoral Immune Responses in Neurosyphilis: CXCL13/CXCR5 Play a Pivotal Role for B-Cell Recruitment to the Cerebrospinal Fluid.

doi: 10.1093/infdis/jix233

Figure Lengend Snippet: Figure 5. Pathologic diagnosis of intracranial syphilitic gumma and cellular organization of follicle-like structures. HE staining of intracranial syphilitic gumma (first row). Immunostaining of spirochete in intracranial syphilitic gumma (second row). Immunostaining of CXCL13 in intracranial syphilitic gumma and follicle-like structure (third row). Immunostaining of CXCR5 in intracranial syphilitic gumma and follicle-like structure (fourth row). Immunostaining of CD20+ B cells in intra- cranial syphilitic gumma and follicle-like structure (fifth row). Immunostaining of CD3+ T cells in intracranial syphilitic gumma and follicle-like structure (sixth row). Immunostaining of CD35+ FDCs in intracranial syphilitic gumma and follicle–like structure (seventh row). Immunostaining of CD138+ plasma cells in intracranial syphilitic gumma and follicle-like structure (eighth row). Abbreviations: FDCs, follic- ular dendritic cells; HE, hematoxylin and eosin.

Article Snippet: Paraffin-embedded brain biopsy (4 μm) was stained with appropriate concentration of primary antibodies against CXCL13, CXCR5, CD20, CD3, CD35, CD138 (Maixin-Bio, Fujian, China), and spirochete (Biocare Medical, Pacheco, CA), then incubated with streptavidin-biotin complex system according to manufacturer’s instructions (Boster, Pleasanton, CA).

Techniques: Biomarker Discovery, Staining, Immunostaining, Clinical Proteomics

Journal: International Journal of Molecular Sciences

Article Title: Low-Dose Salinomycin Alters Mitochondrial Function and Reprograms Global Metabolism in Burkitt Lymphoma

doi: 10.3390/ijms26115125

Figure Lengend Snippet: Salinomycin (SAL) reduces mitochondrial membrane potential (MMP) in live Raji cells. ( A ) MMP was assessed using the cationic carbocyanine dye JC-1 after 12 h of SAL treatment at concentrations of 0.25 µM, 0.5 µM, and 5 µM. ( B ) MMP was evaluated using tetramethylrhodamine methyl ester (TMRM) following 36 h of SAL treatment at the same concentrations. A positive control was included by treating cells with 50 µM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) for 1 h. Data are expressed as mean ± SD from three experiments ( n = 3), and statistical significance was assessed using one-way ANOVA (*** p < 0.001, **** p < 0.0001).

Article Snippet: The Burkitt lymphoma Raji cell line (RRID:CVCL_0511) was maintained in RPMI 1640 medium (Corning, NY, USA; cat. #10-040-CVR) supplemented with 10% fetal bovine serum (FBS; Fisher Scientific, Waltham, MA, USA; cat. #SH3007203), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Membrane, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Low-Dose Salinomycin Alters Mitochondrial Function and Reprograms Global Metabolism in Burkitt Lymphoma

doi: 10.3390/ijms26115125

Figure Lengend Snippet: SAL induces the generation of reactive oxygen species (ROS) in Raji cells. ( A ) Mitochondrial superoxide production was detected using MitoSOX dye following 36 h of SAL treatment at concentrations of 0.25 µM, 0.5 µM, and 5 µM. ( B ) Total ROS levels were assessed using CellROX Green reagent after 48 h of SAL treatment at concentrations of 0.25 µM and 0.5 µM. As a positive control, cells were treated with 50 µM CCCP for 1 h. Results are expressed as mean ± SD from three experiments ( n = 3), and statistical significance was evaluated using one-way ANOVA (**** p < 0.0001).

Article Snippet: The Burkitt lymphoma Raji cell line (RRID:CVCL_0511) was maintained in RPMI 1640 medium (Corning, NY, USA; cat. #10-040-CVR) supplemented with 10% fetal bovine serum (FBS; Fisher Scientific, Waltham, MA, USA; cat. #SH3007203), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Low-Dose Salinomycin Alters Mitochondrial Function and Reprograms Global Metabolism in Burkitt Lymphoma

doi: 10.3390/ijms26115125

Figure Lengend Snippet: SAL suppresses mitochondrial respiration in Raji cells. ( A ) Oxygen consumption rate (OCR) was measured using a Seahorse XFe96 analyzer, following 24 h-long treatment with 0.25 µM, 0.5 µM, or 5 µM SAL. OCR was assessed under basal conditions and after the sequential addition of oligomycin (2.5 µM), carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, 1 µM and 1.5 µM), and antimycin A (2.5 µM). The data concerning maximal respiration ( B ), basal respiration ( C ), and ATP production ( D ) have been extracted from the analysis presented in ( A ). Data were normalized to cell number and represent the mean ± SD from experiments ( n = 3). Statistical analysis was performed using one-way ANOVA (** p < 0.01, **** p < 0.0001).

Article Snippet: The Burkitt lymphoma Raji cell line (RRID:CVCL_0511) was maintained in RPMI 1640 medium (Corning, NY, USA; cat. #10-040-CVR) supplemented with 10% fetal bovine serum (FBS; Fisher Scientific, Waltham, MA, USA; cat. #SH3007203), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Low-Dose Salinomycin Alters Mitochondrial Function and Reprograms Global Metabolism in Burkitt Lymphoma

doi: 10.3390/ijms26115125

Figure Lengend Snippet: SAL reduces glycolytic activity in Raji cells. ( A ) Extracellular acidification rate (ECAR) was measured using the Seahorse XFe96 analyzer following treatment with 0.25 µM, 0.5 µM, and 5 µM SAL under basal conditions and after sequential injections of oligomycin (2.5 µM), FCCP (1 µM and 1.5 µM), and antimycin A (2.5 µM). ( B , C ) Quantification of basal ECAR ( B ) and glycolytic reserve in response to oligomycin ( C ) was extracted from the ECAR profiles shown in ( A ). Data were normalized to cell number and represent the mean ± SD from three experiments ( n = 3). Statistical analysis was performed using one-way ANOVA (** p < 0.01, ns indicates non-significant differences ( p > 0.05)).

Article Snippet: The Burkitt lymphoma Raji cell line (RRID:CVCL_0511) was maintained in RPMI 1640 medium (Corning, NY, USA; cat. #10-040-CVR) supplemented with 10% fetal bovine serum (FBS; Fisher Scientific, Waltham, MA, USA; cat. #SH3007203), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Low-Dose Salinomycin Alters Mitochondrial Function and Reprograms Global Metabolism in Burkitt Lymphoma

doi: 10.3390/ijms26115125

Figure Lengend Snippet: Untargeted metabolomic profiling of Raji cells treated with 0.25 µM SAL, analyzed using SolariX 2xT 7T. ( A ) OPLS-DA (Orthogonal Projections to Latent Structures–Discriminant Analysis) score plot R2X = 0.104, R2Y = 0.878, Q2 = 0.476; CV-ANOVA p = 0.0399. ( B ) OPLS-derived VIP (Variable Importance in Projection) score plot.

Article Snippet: The Burkitt lymphoma Raji cell line (RRID:CVCL_0511) was maintained in RPMI 1640 medium (Corning, NY, USA; cat. #10-040-CVR) supplemented with 10% fetal bovine serum (FBS; Fisher Scientific, Waltham, MA, USA; cat. #SH3007203), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Low-Dose Salinomycin Alters Mitochondrial Function and Reprograms Global Metabolism in Burkitt Lymphoma

doi: 10.3390/ijms26115125

Figure Lengend Snippet: Metabolome of SAL-treated Raji cells. Volcano plots display differentially regulated metabolites, with the most significant ones marked with annotations, based on ( A ) FDR-adjusted p -values ( t -test p = 0.05) and ( B ) unadjusted p -values ( t -test p = 0.05). Blue represents metabolites that are lower in the SAL-treated group, while red denotes metabolites that are higher relative to the vehicle group. Horizontal dashed line indicates p = 0.05.

Article Snippet: The Burkitt lymphoma Raji cell line (RRID:CVCL_0511) was maintained in RPMI 1640 medium (Corning, NY, USA; cat. #10-040-CVR) supplemented with 10% fetal bovine serum (FBS; Fisher Scientific, Waltham, MA, USA; cat. #SH3007203), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.

Techniques: