diffuse large b cell lymphoma dlbcl cell lines su dhl 5  (DSMZ)

Journal: Nature immunology

Article Title: Dynamic regulation of B cell Complement Signaling is Integral to Germinal Center Responses

doi: 10.1038/s41590-021-00926-0

Figure Lengend Snippet: a . Schematic diagram illustrating effects of absent DAF with persistent CD59 expression on complement activation products on naïve (top) vs GC (bottom) B cells. b. Gating strategy for DAF/CD55 expression on immunized BCL6-YFP + reporter mice, d3 post-immunization with SRBCs including d10 naïve and GC B cell controls (complementary to ). c . Heat map depicting relative mRNA expression levels (row-normalized) for BCL6 and complement regulators on human B cell lymphoma cell lines (Diffuse Large B cell lymphoma and selected Burkitt lymphoma cell lines: Raji, BL70, P3HR1; and Multiple Myeloma cell lines: KMS26, KMS27 and MOLP2). Data extracted from the Cancer Cell Line Encyclopedia (CCLE) repository , . ( d-e ) Schematic depiction of ChIP-seq tracks for BCL6 and selected histone marks at the human RCA ( d ) and CD59 ( e ) gene loci. Data extracted from GEO records GSE68349, GSE67494 , .

Article Snippet: The human diffuse large B cell lymphoma (DLBCL) cell lines SU-DHL-5, SU-DHL-6, SU-DHL-10, OCI-LY7, TOLEDO and the Burkitt lymphoma cell line P3HR1, were gifts of L. Pasqualucci (Columbia University Medical Center) and originally purchased from the DSMZ ( www.dsmz.de ) repository (SU-DHL lines and OCI-LY7) or ATCC ( atcc.org ) (P3HR1 and TOLEDO).

Techniques: Expressing, Activation Assay, ChIP-sequencing

Journal: Nature immunology

Article Title: Dynamic regulation of B cell Complement Signaling is Integral to Germinal Center Responses

doi: 10.1038/s41590-021-00926-0

Figure Lengend Snippet: a. Representative histogram (left) and quantified results (right) of DAF (CD55) and Bcl-6 on BCL-6-YFP + B cells, d3 post-immunization (gating strategy, ); mean±SEM. b. ChIPseq analysis of human GC B cells (tonsil) showing distribution of BCL-6, H3K4Me1 and H3K4Me3 marks at the CD55 gene (from GSE68349, GSE67494) , . c-d. Representative histograms for BCL-6 ( c , left) and DAF ( c , right) with quantification ( d ) for BCL-6, DAF, CD46 and CD59 on KMS27 (multiple myeloma) cells following BCL-6 overexpression (n=5 for BCL-6 and DAF, n=3 for CD46; n=2 for CD59). e-f . Quantified changes in DAF, CD46 and CD59 expression on SUDHL5 DLBCL B-lymphoma cells ( e ), (DAF & CD59, n=3; CD46, n=6); following transfection with a dominant negative (ZF) BCL-6 ( f , representative immunoblot). The anti-BCL-6 antibody recognizes a region present in the ZF domain of this protein (BCL-6, full-length protein) g-i. Fold change in surface expression of DAF ( g ), CD46 ( h ) and CD59 ( i ) by flow cytometry 24h after addition of BCL-6 inhibitor FX1 (50 mM), or DMSO control. FX1 significantly upregulated DAF in 5 cell lines (SUDHL5, n=6; SUDHL6, n=4; SUDHL10, n=8; P3HR1, n=2; OCI-LY7, n=3). p value is a summary of all replicates for all lines. FX1 upregulated CD46 in OCI-LY7, n=1 and SUDHL10, n=2 (p value shown for these 3 combined replicates) without effect in SUDHL5, n=2 and P3HR1 n=2 (p value for these combined replicates: ns). FX1 upregulated CD59 in 3/3 lines (SUDHL5, n=3; SUDHL6, n=4; SUDHL10 n=3). p value is shown for these 10 combined replicates from the 3 cell lines. Data are presented as normalized to DMSO levels for each experiment j . left: construct schematics for BCL-6, BCL-6 DNA binding domain ZF and BCL-6 that lacks DNA binding domain delta ZF. right: quantified luciferase signal under each condition, For all panels, data are presented as mean values +/− SEM, *p<0.05, **p<0.01, ***p<0.005, ****p<0.001 by ANOVA with Bonferroni post-test ( a, j ) unpaired t-test, two-tailed ( a,d-e ) or paired t-test, two-tailed ( g ). ns, not significant.

Article Snippet: The human diffuse large B cell lymphoma (DLBCL) cell lines SU-DHL-5, SU-DHL-6, SU-DHL-10, OCI-LY7, TOLEDO and the Burkitt lymphoma cell line P3HR1, were gifts of L. Pasqualucci (Columbia University Medical Center) and originally purchased from the DSMZ ( www.dsmz.de ) repository (SU-DHL lines and OCI-LY7) or ATCC ( atcc.org ) (P3HR1 and TOLEDO).

Techniques: Over Expression, Expressing, Transfection, Dominant Negative Mutation, Western Blot, Flow Cytometry, Control, Construct, Binding Assay, Luciferase, Two Tailed Test

Journal: Human Molecular Genetics

Article Title: XIAP and cIAP1 amplifications induce Beclin 1-dependent autophagy through NFκB activation

doi: 10.1093/hmg/ddv052

Figure Lengend Snippet: High levels of XIAP in B-cell lymphoma activate autophagy. ( A ) B cells (WT) and three diffuse large B-cell lymphoma cell lines, SUDHL5, SUDHL8 and SUDHL10 were subjected to immunoblotting with XIAP and actin antibodies. ( B ) The same cell lines were treated with 400 n m bafilomycin A1 and were subsequently subjected to immunoblotting with LC3 and tubulin antibodies. The diffuse large B-cell lymphoma cell lines, SUDHL5 ( C ), SUDHL8 ( D ) and SUDHL10 ( E ), were treated without or with 10 μM embelin (Emb) for 16 h, and treated without or with 400 n m bafilomycin A1 (Baf A1) for the last 4 h of the experiment, and were subsequently subjected to immunoblotting with LC3 and tubulin antibodies. ( F ) Propidium iodide and FITC-conjugated Annexin A5 staining detected by flow cytometry of B cells (WT) SUDHL5, SUDHL8 and SUDHL10 (5, 8 and 10) treated without or with 10 μM embelin (Emb) for 16 h. The percentage of cells positive for both PI and Annexin A5 are shown. Densitometric measurements of LC3-II bands were normalized to the corresponding actin or tubulin bands and are shown in the corresponding histograms. The values shown in all the histograms represent the mean ± standard deviation from at least three independent experiments performed in triplicate samples/condition. The P -values were determined using one sample t -tests, where controls are set to 100%. See also Supplementary Material, Figure S7 .

Article Snippet: Wild-type B-cells and human diffuse large B-cell lymphoma cell lines (DLBCL) SUDHL5, SUDHL8 and SUDHL10 [obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany (DSMZ)] were cultured at 37°C, 5% CO 2 in 10% FBS, 2 m m l -glutamine and 100 U/ml penicillin/streptomycin supplemented RPMI 1640 (Invitrogen) (see Supplementary Material online ).

Techniques: Western Blot, Staining, Flow Cytometry, Standard Deviation