Journal: bioRxiv

Article Title: CRISPR Screens Reveal Epstein-Barr Virus-activated JunB as a Key Lymphoblastoid B cell Dependency Factor that Represses Cyclin Dependent Kinase Inhibitor P18INK4c

doi: 10.64898/2026.03.31.715635

Figure Lengend Snippet: (A) Log2(fold change) of the four guides targeting JunB (red) compared to the distribution of all sgRNA guides from the GM12878 and P3HR-1 Avana dropout screen. Fold change was calculated comparing abundance of sgRNA day 21 post selection versus input. (B) JunB sgRNA abundances in Brunello and Avana dropout screens. Log 2 JunB sgRNA abundances from GM12878 (red) and P3HR-1 (blue) cells 21 days post library selection were determined via deep sequencing and compared to input library sgRNA abundance (green). (C) c-Jun sgRNA abundances in Brunello and Avana dropout screens. Log 2 c-Jun sgRNA abundances from GM12878 (red) and P3HR-1 (blue) cells 21 days post library selection were determined via deep sequencing and compared to input library sgRNA abundance (green). (D) JunD sgRNA abundances in Brunello and Avana dropout screens. Log 2 JunD sgRNA abundances from GM12878 (red) and P3HR-1 (blue) cells 21 days post library selection were determined via deep sequencing and compared to input library sgRNA abundance (green). (E) JunB dependency across CRISPR DepMap lymphoid cell lines ( DepMap.org ). Gene dependency scores were calculated by comparing JunB sgRNA abundance in individual lymphoid cell lines 21 days post lentivirus library transduction and selection in comparison to input library abundance. Each circle indicates an individual lymphoid cell line. Circle size represents Log JunB reads from RNA-seq analysis (DepMap, Broad, DepMap Public 25Q3 (2025)) Green circles versus clear circles indicate DepMap Burkitt versus non-Burkitt lymphoid cell lines, respectively. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005

Article Snippet: Antibodies against the following proteins were used in this study: JunB (Cell Signaling Technology, #3753), c-Jun (Cell Signaling Biotechnology, #9165), JunD (Cell Signaling Biotechnology, #5000), β-Actin (Biolegend, #664802), LMP1 (Abcam, ab78113), LMP2A (Abcam, ab59028), EBNA2 PE2 (a gift from Fred Wang), DDX1 (Bethyl, A300-521A-M), Myc (Santa Cruz Biotechnology, SC-40), p100/p52 (EMD Millipore, 05-361), TRAF1 (Cell Signaling Biotechnology, #4715S), HA tag antibody (Cell Signaling Technology, # 3724), BIM (Cell Signaling Biotechnology, #2933), CCND2 (Proteintech, 10934-1-AP), EBNA1 (a gift from Jaap Middeldorp), and EBNA3C A10 (hybridoma from Martin Rowe’s group[ ]).

Techniques: Selection, Sequencing, CRISPR, Transduction, Comparison, RNA Sequencing

Journal: bioRxiv

Article Title: CRISPR Screens Reveal Epstein-Barr Virus-activated JunB as a Key Lymphoblastoid B cell Dependency Factor that Represses Cyclin Dependent Kinase Inhibitor P18INK4c

doi: 10.64898/2026.03.31.715635

Figure Lengend Snippet: (A) Validation of JunB CRISPR-Cas9 knockout in MUTU I and GM15892 cells. Immunoblot was performed on WCL from Cas9+ MUTU I and GM15892 cells transduced with control or JunB sgRNA seven days post-selection using indicated antibodies. (B) Mean ± SD live cell numbers of Cas9+ MUTU I or GM11830 cells expressing control or JunB targeting single guide RNAs (sgRNA) from n=3 replicates. Cells transduced with lentiviruses expressing the indicated sgRNAs were puromycin selected. Cell numbers two days following puromycin selection (defined as day 0 of the graph) were set to 1. Live cell numbers were quantitated by CellTiter-Glo assay (C) Investigation of impact of JunB on other Jun AP-1 transcription factors. Immunoblot analysis of WCL from Cas9+ GM12878 cells transduced with control or JunB sgRNAs, ten days post selection, with indicated antibodies. Densitometry ratios of JunB and JunD to β-Actin is shown. (D) Investigation of combinatorial impact of JunB and JunD on LCL growth. (Left) Mean ± SD live cell numbers of Cas9+ GM12878 cells expressing control, JunD, JunB, or JunB+JunD targeting single guide RNAs (sgRNA) from n=3 replicates. Cells transduced with lentiviruses expressing control or JunD sgRNA were Zeocin selected two days post transduction. After one week, cells were then transduced with a separate control or JunB sgRNA. Cell numbers two days following puromycin selection (defined as day 0 of the graph) were set to 1. Live cell numbers were quantitated by CellTiter-Glo assay. (Right) Immunoblot validation of JunB and JunD knockouts. WCL of GM12878 cells transduced with both zeocin and puromycin sgRNAs was extracted two days following puromycin selection and immunoblot was performed using indicate antibodies. (E) PI cell cycle stainin of GM12878 cells transduced with JunB sgRNA. Representative FACS cell cycle plots from n=3 replicates of GM12878 cells analyzed ten days post selection. P-values were determined by one-sided Fisher’s exact test. * p<0.05, **p<0.005, ***p<0.0005

Article Snippet: Antibodies against the following proteins were used in this study: JunB (Cell Signaling Technology, #3753), c-Jun (Cell Signaling Biotechnology, #9165), JunD (Cell Signaling Biotechnology, #5000), β-Actin (Biolegend, #664802), LMP1 (Abcam, ab78113), LMP2A (Abcam, ab59028), EBNA2 PE2 (a gift from Fred Wang), DDX1 (Bethyl, A300-521A-M), Myc (Santa Cruz Biotechnology, SC-40), p100/p52 (EMD Millipore, 05-361), TRAF1 (Cell Signaling Biotechnology, #4715S), HA tag antibody (Cell Signaling Technology, # 3724), BIM (Cell Signaling Biotechnology, #2933), CCND2 (Proteintech, 10934-1-AP), EBNA1 (a gift from Jaap Middeldorp), and EBNA3C A10 (hybridoma from Martin Rowe’s group[ ]).

Techniques: Biomarker Discovery, CRISPR, Knock-Out, Western Blot, Transduction, Control, Selection, Expressing, Glo Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Fibroblast growth factor-inducible 14 accelerates pulmonary fibrosis by inducing fibroblast senescence in mice

doi: 10.1007/s00018-026-06161-w

Figure Lengend Snippet: Fn14 shRNA alleviates BLM-induced PF in mice. A , C57BL/6J mice received empty adenovirus or Fn14 vectors (5 × 10 8 PFU/20 g, i.t. ). Seven days later, mice were administered BLM (2.5 mg/kg, i.t. ) to establish a PF model. Mice were sacrificed after the BLM administration on the 21st day. B , the expression of Fn14 mRNA in the lungs was detected by Real-time PCR. C-D , the expression of Fn14 protein in the lungs was detected by Western blot ( n = 6–8). E, weight change of mice ( n = 6–10). F, the rate of weight change in mice was calculated. G, the pulmonary tissue histopathology of mice was stained with H&E (Top row), and the collagen deposition was detected by Masson’s trichrome staining (Bottom row). H, the Ashcroft score was evaluated by three blinded pathologists. I, the collagen subtype was detected by Sirius red staining. J, the expressions of Col1a1 , Col3a1 , and Acta2 mRNA in the lungs were detected by Real-time PCR. L, the expressions of Collagen I, Collagen III, and α-SMA proteins in the lungs were assayed by Western blot ( n = 6–8). ***P < 0.001

Article Snippet: Anti-α-SMA polyclonal antibody , CST , 19,245 , 1: 5000.

Techniques: shRNA, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Histopathology, Staining