sudhl8  (DSMZ)

Journal: Cell Communication and Signaling : CCS

Article Title: GALNT3 is a novel target driving lymphomagenesis via O-glycosylation of FGFR2

doi: 10.1186/s12964-025-02599-w

Figure Lengend Snippet: GALNT3 sustains B cell lymphomagenesis by enhancing DLBCL proliferation. Total proteins were extracted from a panel of DLBCL cells prior to subject either to ( A ) qPCR or ( B ) Immunoblotting of GALNT3 expression levels. GALNT3 low expressing cells (Karpas-422, DB and SUDHL8) were transduced with lentiviral encoding empty vector or GALNT3 overexpression plasmids for 48 h, followed by flow cytometry sorting of GFP + cells to obtain stable transfectants. GALNT3 overexpression efficiency was determined by ( C ) qPCR or ( D ) Immunoblotting. Graph with error bars represent the mean ± SD from technical triplicates (* p < 0.01; ** p < 0.05; *** p < 0.001) compare empty vector to GALNT3 overexpression by Two-way ANOVA with replication . E CFSE assay of DLBCL cell proliferation, ( F ) Cell countess analysis of cell growth, ( G ) Brdu labelling of cell cycle, quantitation of percentage of cell cycle distribution was determined by gating of live cells in either G1 (light blue bar), S (light red bar) or G2/M (light green bar) phases. Graph with error bars represent the mean + SD from technical triplicates (* p < 0.01; ** p < 0.05; *** p < 0.001) compare empty vector to GALNT3 overexpression between G1 phase and S phase by Two-way ANOVA with replication. H above stable transfectants were synchronized to G1/S phase either by double thymidine or serum starvation methods accordingly to a published protocol followed by Immunoblotting assay to determine the expressions of cell cycle regulated proteins. a tubulin was included as an indication of equal loading of total amount of proteins per lane. All experiments were repeated three times. Graph with error bars represent the mean ± SD from technical triplicates (* p < 0.01; ** p < 0.05; *** p < 0.001) compare empty vector to GALNT3 overexpression by unpaired student T test

Article Snippet: SUDHL8 (CRL-2961), and DB (CRL-2289) cells were obtained from the American Type Culture Collection (ATCC), while Karpas-422 (ACC 32) and Riva (ACC 585) cells were purchased from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbHCell lines (DSMZ).

Techniques: Western Blot, Expressing, Transduction, Plasmid Preparation, Over Expression, Flow Cytometry, CFSE Assay, Quantitation Assay

Journal: Cell Communication and Signaling : CCS

Article Title: GALNT3 is a novel target driving lymphomagenesis via O-glycosylation of FGFR2

doi: 10.1186/s12964-025-02599-w

Figure Lengend Snippet: GALNT3 upregulates FGFR2 mediated MAPK signaling pathway in DLBCL. Total RNA was extracted from either GALNT3 overexpressed Karpas-422 or SUDHL8 stable transfectants, RNA library was prepared using KAPA Stranded RNA-SEQ Library Prep Kit. A & B KEGG analysis of GALNT3 responsive genes categorized top 10 signaling pathways ( C ) Gene sets enrichment of GALNT3-responsive genes shows up regulation of the FGFR2 mediated MAPK signaling pathway in either GALNT3 overexpressed Karpas-422 or SUDHL8 cells vs. empty vector, ( D - E ) qPCR validation of fold changes of FGFR gene expressions in lentiviral encoding PCDH-GALNT3 transduced Karpas-422 and SUDHL8 cells. F - G qPCR validation of fold changes of MAPK gene expressions in lentiviral encoding PCDH-GALNT3 transduced Karpas-422 and SUDHL8 cells. * denote FGFR2 signaling and MAPK signaling pathways. All experiments were repeated three times. Graph with error bars represent the mean ± SD from technical triplicates (* p < 0.01; ** p < 0.05; *** p < 0.001, **** p < 0.005) compare empty vector to GALNT3 overexpression by unpaired student T test

Article Snippet: SUDHL8 (CRL-2961), and DB (CRL-2289) cells were obtained from the American Type Culture Collection (ATCC), while Karpas-422 (ACC 32) and Riva (ACC 585) cells were purchased from Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbHCell lines (DSMZ).

Techniques: RNA Sequencing, Protein-Protein interactions, Plasmid Preparation, Biomarker Discovery, Over Expression