hodgkin lymphoma-derived cell line l428  (Creative Bioarray Inc)

Journal: bioRxiv

Article Title: Genome-scale spatial mapping of the Hodgkin lymphoma microenvironment identifies tumor cell survival factors

doi: 10.1101/2025.01.24.631210

Figure Lengend Snippet: a , Barplot showing the effect of recombinant factors on the relative viability (normalized to untreated control) of L1236 cells at three days under low serum conditions; mean ± s.d. is depicted (n = 3); P < 0.05 is considered significant; two-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test. b , waterfall plots (ordered by increasing CRISPR gene effect score) showing genetic dependencies of 3 cHL cell lines (L1236, UHO1, L428). c, d . scatterplot of IL4R and IL13RA1 (c) and IL4R and STAT6 (d) gene effect scores across the DepMap dataset. e , effect of CRISPR/Cas9-mediated knock out of IL13RA1 or IL4R on L1236 cell viability (viable cell counts). f , rainplot of IL13 expression showing that the cHL cell lines (L1236, L428, and UHO1) show outlier expression of IL13. g , effect of CRISPR/Cas9-mediated knock out of IL13 on L1236 cell viability (viable cell counts), with or without recombinant IL13 supplementation (rIL13). h , dose-response analysis of tralokinumab, lebrikizumab, and dupilumab on L1236 cells. IC 50 values (95% confidence interval) are indicated on the right (n = 2); mean ± s.d. is depicted. The relative viability values represent the mean of two independent experimental replicates. i , time course analysis of the effect of tralokinumab, lebrikizumab, dupilumab (0.5 µM each), and isotype control antibody on L1236 cell viability (n = 3); mean ± s.d. is depicted. The C max range of these antibodies reported in clinical trials is indicated in grey. j , schematic representation of the proposed model of spatial organization and interactions that mediate tumor cell growth within the intact cHL tumor microenvironment. The data in e, g, and i are representative of three independent experiments.

Article Snippet: Mammalian cell lines L428, L1236, and UHO1 were obtained from the Leibniz Institute DSMZ and cultured in media and conditions recommended by the distributor.

Techniques: Recombinant, Control, Comparison, CRISPR, Knock-Out, Expressing, Clinical Proteomics

Journal: Oncogenesis

Article Title: CCND3 is indispensable for the maintenance of B-cell acute lymphoblastic leukemia

doi: 10.1038/s41389-021-00377-0

Figure Lengend Snippet: A The mRNA expression data of CCND1 , CCND2 , and CCND3 in different B-ALL subtypes were mined from publicly available databases using the GENEVESTIGATOR software ( http://www.genevestigator.com/ ). The experiment IDs are listed in Supplementary Materials and Methods. Data shown as mean ± SD. B The mRNA expression levels of D-type cyclins in B–ALL (blue) and control cell lines was measured by qRT-PCR. The control cell lines included Burkitt lymphoma (Ramos), cHL (L428), and HEK293T which express high levels of CCND3 , CCND2 , and CCND1 , respectively. Data shown as mean ± SD, n = 3. C Ratio of mRNA levels CCND3/CCND2 measured by qRT-PCR in B-ALL cell lines. D Expression of CCND2 and CCND3 proteins in B-ALL cell lines was measured by immunoblot. TUBB is used as loading control. Image representative of n = 3.

Article Snippet: B-ALL cell lines NALM-6, RS4;11, BV-173, REH, TOM-1, NALM-20, SUP-B15, KOPN-8, and EU-3/697, the classical Hodgkin lymphoma (cHL) cell line L428 and Burkitt lymphoma cell line RAMOS, HEK293T, and LentiX cells were purchased from DSMZ (Braunschweig, Germany).

Techniques: Expressing, Software, Control, Quantitative RT-PCR, Western Blot

Journal: Cancer Science

Article Title: Epstein‐Barr virus latent membrane protein‐1 upregulates autophagy and promotes viability in Hodgkin lymphoma: Implications for targeted therapy

doi: 10.1111/cas.14833

Figure Lengend Snippet: LMP1 increases autophagy in HL cells and helps HL cells adapt to starvation‐induced autophagic stress. A, Expression patterns of LMP1 in LCL, parental EBV‐negative L428 or KM‐H2 cell lines. The cell lines (LCL, L428, and KM‐H2) were blotted with antibodies against LMP1 and GAPDH. GAPDH served as the loading control. B, Expression of LMP1 and autophagic markers in PBMCs infected with EBV. The samples from parental PBMCs, EBV‐infected newly formed (#1) and long‐termed (#2) LCLs were blotted with αLMP1 (LMP1) and αLC3 (LC3‐I and ‐II) antibodies. The red numbers represent relative band intensities normalized by GAPDH. C, Expression of autophagic markers in L428‐LMP1 (left) and KM‐H2‐EBV (right) cells. C, left panel, The LMP1‐transfected L428 cells had undergone stable selection and show expression of LMP1 but not EBNA1 or EBNA2. Stably transfected L428 cells with a GFP‐expressing vector served as the nonexpressing control. LMP1 transfection increased LC3‐II expression but decreased expression of other autophagic proteins. C, right panel, KM‐H2‐EBV cells expressed EBNA1 but not LMP1 or EBNA2. Stably‐transfected KM‐H2 cells with a GFP‐expressing vector served as the nonexpressing control. EBV type I latency program augmented Atg7 expression but attenuated expression of LC3‐II, Atg5 and Beclin 1. The red numbers represent the relative band intensity of LC3‐II normalized by GAPDH. D, The changes in autophagic markers induced by starvation in L428 HL cells. The L428‐GFP and L428‐LMP1 cells (each 2 × 10 5 cells) were cultured in 100 μL of 2.5% FBS medium for 72 h. The samples were harvested at respective time points and assayed by blotting with antibodies against Beclin1, Atg7, Atg5, p62, LC3, and GAPDH. The numbers represent the relative band intensities normalized by GAPDH (red for LC3‐II). The changes of LC3‐II ratio (starvation divided by nonstarvation) are plotted. Other autophagic proteins show a similar trend with LC3‐II. E, Stable cell lines (L428‐GFP and L428‐LMP1) were stained by αLC3 antibody on immunofluorescence, which shows more LC3‐II signals (red) in L428‐LMP than those in L428‐GFP cells (31/383 = 8.1% vs 9/449 = 2.0%, P < .001; blue, DAPI for nuclear staining)

Article Snippet: The EBV‐negative HL cell lines L428 and KM‐H2 (DSMZ) were grown in RPMI‐1640 medium supplemented with 10% fetal bovine serum (HyClone).

Techniques: Expressing, Control, Infection, Transfection, Selection, Stable Transfection, Plasmid Preparation, Cell Culture, Staining, Immunofluorescence

Journal: Cancer Science

Article Title: Epstein‐Barr virus latent membrane protein‐1 upregulates autophagy and promotes viability in Hodgkin lymphoma: Implications for targeted therapy

doi: 10.1111/cas.14833

Figure Lengend Snippet: LMP1 increases autophagic flux LC3‐II and rescues chloroquine (CQ)‐induced death in HL cells. L428‐LMP1 and the control L428‐GFP (1.5 × 10 6 cells for each) were treated with 0, 5, or 10 µM chloroquine (CQ) for 24 h (A), 48 h (B), and 72 h. C, left panel, The samples were harvested and blotted with αLMP1, αBeclin1, αAtg7, αAtg5, αp62, αLC3, and αGAPDH antibodies. Red numbers indicate relative band intensities of LC3‐II normalized by GAPDH. Other autophagic proteins show a similar pattern with LC3‐II or unremarkable changes in protein expression. C, middle panel, Representative flow‐cytometry plots to evaluate the apoptotic cell death occurring in the identical samples in the left panel, which were harvested and stained for Annexin V (FL2) and 7‐AAD (FL3). C, right panel, Quantification of the flow data in the middle panel. The percentage of cells in quadrants 2 + 3 was used for plotting (n = 3, mean ± SEM). Asterisks represent P < .05 by paired t ‐test (* P < .05, ** P < .01, *** P < .001). D, LC3‐II ratios are plotted based on relative band intensities of LC3‐II normalized by GAPDH (red numbers)

Article Snippet: The EBV‐negative HL cell lines L428 and KM‐H2 (DSMZ) were grown in RPMI‐1640 medium supplemented with 10% fetal bovine serum (HyClone).

Techniques: Control, Expressing, Flow Cytometry, Staining

Journal: Cancer Science

Article Title: Epstein‐Barr virus latent membrane protein‐1 upregulates autophagy and promotes viability in Hodgkin lymphoma: Implications for targeted therapy

doi: 10.1111/cas.14833

Figure Lengend Snippet: LMP1 increases autophagic flux LC3‐II and rescues doxorubicin (DOX)‐induced death in HL cells. L428‐LMP1 and the control L428‐GFP (1.5 × 10 6 cells for each) were treated with 0, 0.1, 1, or 10 µM doxorubicin (DOX) for 24 h (A), 48 h (B), and 72 h. C, left panel, The samples were harvested and blotted with αBeclin1, αAtg7, αAtg5, αp62, αLC3 and αGAPDH antibodies. Red numbers indicate relative band intensities of LC3‐II normalized by GAPDH. In contrast to LC3‐I/II, other autophagic proteins demonstrated decreased expression along with HL cell death, especially at 10 μM DOX and at 48 and 72 h. C, middle panel, Representative flow‐cytometry plots to evaluate the apoptotic cell death occurring in the identical samples in the left panel, which were harvested and stained for Annexin V (FL2) and 7‐AAD (FL3). C, right panel, Quantification of the flow data in the middle panel. The percentage of cells in quadrants 2 + 3 was used for plotting (n = 3, mean ± SEM). Asterisks represent P value <.05 by paired t ‐test (* P < .05, ** P < .01, *** P < .001). D, Plots of the LC3‐II ratio normalized by GAPDH in immunoblotting

Article Snippet: The EBV‐negative HL cell lines L428 and KM‐H2 (DSMZ) were grown in RPMI‐1640 medium supplemented with 10% fetal bovine serum (HyClone).

Techniques: Control, Expressing, Flow Cytometry, Staining, Western Blot

Journal: Cancer Science

Article Title: Epstein‐Barr virus latent membrane protein‐1 upregulates autophagy and promotes viability in Hodgkin lymphoma: Implications for targeted therapy

doi: 10.1111/cas.14833

Figure Lengend Snippet: Xenograft mouse model shows that CQ treatment effectively eradicated L428‐LMP1 xenograft more efficiently than did L428‐GFP xenograft. NOD/SCID mice were divided into four groups: GFP + PBS (n = 6), GFP + CQ (n = 7), LMP1 + PBS (n = 7), and LMP1 + CQ (n = 7). GFP, L428‐GFP xenograft; LMP1, L428‐LMP1 xenograft; CQ, CQ treatment (60 mg/kg for 25 days); PBS, PBS mock treatment control. A, Appearances of the mice and their injection sites on the back 25 days after tumor cell injection and treatment. B, The gross morphology of the excised tumors in (A). C‐F, Plots of the xenograft tumor sizes (C, mm 3 ; D, percentages of the original size), tumor weights (E, grams), and the corresponding mouse weights (F, grams). Black, GFP + PBS; green, GFP + CQ; red, LMP1 + PBS; blue, LMP1 + CQ. Asterisks represent P value <.05 by Student t ‐test

Article Snippet: The EBV‐negative HL cell lines L428 and KM‐H2 (DSMZ) were grown in RPMI‐1640 medium supplemented with 10% fetal bovine serum (HyClone).

Techniques: Control, Injection