Journal: Nucleic Acids Research

Article Title: A viral noncoding RNA is a master regulator of gene expression that defines host cell identity and function

doi: 10.1093/nar/gkag472

Figure Lengend Snippet: HSUR1 determines cell identity and function. ( A ) Western blot of representative HSUR1 target proteins in cj38637-WT and cj38637-ΔHSUR1 cells; cofilin serves as a loading control. ( B ) IFN-γ production in cj38637-WT and cj38637-ΔHSUR1 cells detected by western blot after PMA/ionomycin stimulation in the presence of brefeldin A and monensin; quantification of three replicates shown at right. HDAC1 provides a loading control. ( C ) Predicted composition of resting (R) and active (A) immune cell types inferred from gene expression using CIBERSORTx. T-reg, regulatory T cell; γδ-T, gamma-delta T cell; Tfh, T follicular helper cell; DC, dendritic cell; MΦ, macrophage; NK, natural killer cell. ( D ) NK-like cytotoxic activity of WT and ΔHSUR1 cells toward MOLT-4 target cells measured by CytoTox-Glo assay. ( E ) Flow cytometry analysis of cj38637-WT and cj38637-ΔHSUR1 cells for the indicated surface markers. ( F ) Same as in panel (E) for the indicated markers in HuT78 cells (Control) or same cells transduced with a lentiviral vector expressing HSUR1. * P < .05, ** P < 0.01, **** P < .0001.

Article Snippet: MOLT-4 cells (obtained from ATCC, CRL1582) and human HuT78 Sézary Syndrome, cutaneous T lymphocytes (obtained from Vicente Planelles) were grown in RPMI-1640 medium supplemented with 10% FBS, antibiotics, glutamax, and sodium pyruvate.

Techniques: Western Blot, Control, Gene Expression, Activity Assay, Glo Assay, Flow Cytometry, Transduction, Plasmid Preparation, Expressing

Journal: iScience

Article Title: Activation-gated, T cell-restricted silencing of Dapk1 enhances Bacille Calmette-Guérin-elicited protective CD4 + memory

doi: 10.1016/j.isci.2026.115593

Figure Lengend Snippet: Memory phenotype CD4 + T cells downregulate DAPK1 (A and B) Dapk1 is transcriptionally downregulated in CD44 hi CD4 + T cells and converges on apoptotic pathways. (A) Volcano plot displaying DEGs (CD44 hi vs. CD44 lo ). Key apoptosis-related genes are labeled with log 2 FC and p values. (B) Venn diagram illustrating that Dapk1 is uniquely shared across three apoptotic pathways. (C) Dapk1 mRNA expression was quantified by RT-qPCR in CD44 lo , CD44 int , and CD44 hi CD4 + T cell fractions and normalized to Gapdh ( n = 5). (D) Validation of DAPK1 knockdown by RT-qPCR (upper side) and immunoblotting (lower side) in Jurkat cells stably expressing sh DAPK1 or a scramble control ( n = 3). (E) FCM quantification of intracellular active caspase-3 (upper side) and cell viability (using FVS780 dye, lower side) in Jurkat cells 24 h after activation with anti-CD3/CD28 antibodies ( n = 6). (F) Doxycycline-inducible knockdown of DAPK1. DAPK1 mRNA and protein levels were assessed by RT-qPCR and immunoblotting, respectively, in MOLT-4 cells stably expressing a Dox-inducible sh DAPK1 construct 48 h after Dox treatment ( n = 3). (G) FCM quantification of cell viability in the same MOLT-4 cells as in (F) ( n = 3). Data are presented as the mean ± SD. Comparisons between two groups were analyzed using unpaired two-tailed Student’s t test. Comparisons among three or more groups were analyzed using one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Jurket cells MOLT-4 , ATCC ATCC , Cat# TIB-152 Cat# CRL-1582.

Techniques: Labeling, Expressing, Quantitative RT-PCR, Biomarker Discovery, Knockdown, Western Blot, Stable Transfection, Control, Activation Assay, Construct, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Biosensor-guided discovery of peptide inhibitors targeting the ribosomal protein uS5-PDCD2 chaperone interaction

doi: 10.1016/j.jbc.2026.111415

Figure Lengend Snippet: uS5-derived peptides inhibit cancer cell growth. A , Amino acid sequences of wild-type (WT) and F25A/F29A mutant (MT) CPP-uS5 (22–32) peptides, with an N-terminal FITC fluorophore. B , fluorescence microscopy images of HeLa cells incubated with 20 μM WT or MT CPP-uS5 (22–32) peptides. Cells were fixed 24 h post-incubation and analyzed by direct FITC fluorescence (panels b and e). Nuclei were counterstained with DAPI (panels a and d). Scale bars, 20 μm. C-D , Dose–response analysis of cell viability in HeLa ( C ) and MOLT-4 ( D ) cells following 24 h of peptide treatment. Data represent mean ± SD from three independent experiments. E , Half-maximal inhibitory concentration (IC 50 ) values were determined by fitting dose-response curves using nonlinear regression analysis. Statistical differences were calculated using student t-tests. p -values are indicated.

Article Snippet: Human MOLT-4, HEK293T, HeLa, and U2OS cell lines were acquired from ATCC.

Techniques: Derivative Assay, Mutagenesis, Fluorescence, Microscopy, Incubation, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Biosensor-guided discovery of peptide inhibitors targeting the ribosomal protein uS5-PDCD2 chaperone interaction

doi: 10.1016/j.jbc.2026.111415

Figure Lengend Snippet: Inhibition of the uS5-PDCD2 interaction using residues 21 to 50 of uS5. A , Western blot analysis of total extracts (Input, Top ) and anti-Flag purifications (IP:Flag, Bottom ) prepared from HeLa cells that stably expressed Flag-PDCD2 and that were previously transfected with constructs encoding either GFP alone (lane 1), wild-type (WT, lane 2) or F29Y (MT, lane 3) versions of uS5 21-50 -GFP. The blots were analyzed for GFP, Actin, uS5, and Flag-PDCD2. B , quantification of relative uS5 levels copurified from anti-Flag precipitates normalized to Flag-PDCD2 levels. Values were expressed relative to the GFP control vector, which was set to 1.0. The data and error bars represent the average and SD from at least three independent experiments. p -value is indicated, as determined by a one-way ANOVA with Dunnett’s multiple comparisons test. ( C ) U2OS cells that conditionally express Flag-PDCD2 were transfected with either GFP control vector (panels a-d), wild-type (panels e-h) or F29Y (panels i-l) versions of uS5 21-50 -GFP. At the time of transfection, doxycycline was added to the media to induce the expression of Flag-PDCD2. 48 h post-transfection, cells were fixed and simultaneously analysed by direct fluorescence (b, f and j) and immunostaining for Flag-PDCD2 (c, g and k). DNA staining with DAPI shows the nucleus of each cell (a, e and i). Scale bars correspond to 20 μm. D , quantification of nucleus-to-cytoplasmic ratios of Flag-PDCD2 shows a significant decrease in cells that expressed the wild-type version of uS5 21-50 -GFP. More than 60 cells were analyzed for each condition from three independent immunofluorescence experiments. Statistical differences were calculated using a one-way ANOVA with Dunnett’s multiple comparisons test. p -value is indicated. E , Peptide sequences corresponding to residues 21 to 50 of uS5, showing conserved phenylalanines in blue (WT) and substitutions to alanine residues in the mutant in red (MT). F , schematic of the peptide competition experiments using purified uS5-GFP/PDCD2 complex. See text for details. G , Western blot analysis of an uS5-GFP immunoprecipitate that were washed, divided, and treated with increasing concentrations of either wild-type ( Top , WT) or mutant ( Bottom , MT) uS5 21-50 peptides (lanes 2–5) or with no peptide (lane 1). Blots were analyzed for uS5-GFP and endogenous PDCD2. ( H ) Quantification of PDCD2 levels copurified from anti-GFP precipitates normalized to uS5-GFP levels. The values were then set to 1.0 for the control purification in the absence of peptide. Solid lines mark the average binding from three independent replicates, with error bars corresponding to standard deviations. Statistical differences were calculated using a one-way ANOVA with Dunnett’s multiple comparisons test. p -value is indicated.

Article Snippet: Human MOLT-4, HEK293T, HeLa, and U2OS cell lines were acquired from ATCC.

Techniques: Inhibition, Western Blot, Stable Transfection, Transfection, Construct, Control, Plasmid Preparation, Expressing, Fluorescence, Immunostaining, Staining, Immunofluorescence, Mutagenesis, Purification, Binding Assay

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Enzymatic Laccase Nanoreactors Induce Apoptosis in MOLT-4-ALL Cells and Activate Prodrugs in a Synergetic Effect

doi: 10.2147/BLCTT.S576292

Figure Lengend Snippet: Curve dose-response on MOLT-4 leukemia model cells. ( A ) Pro-drugs and ( B ) laccase from C. gallica .

Article Snippet: The MOLT-4 cell line, derived from human T cells originating from acute lymphoblastic leukemia, was obtained from a certified cell bank (ATCC, MOLT-4-CRL-1582).

Techniques:

Journal: Blood and Lymphatic Cancer: Targets and Therapy

Article Title: Enzymatic Laccase Nanoreactors Induce Apoptosis in MOLT-4-ALL Cells and Activate Prodrugs in a Synergetic Effect

doi: 10.2147/BLCTT.S576292

Figure Lengend Snippet: Flow cytometry analysis to determine cell viability. The green zone indicates the death cells with a stain-positive PI, and the red zone indicates the alive cell with a stain-positive FDA. Every panel corresponds to a different treatment of MOLT-4-ALL cell: ( A ) Doxorubicin, ( B ) Doxorubicin + Lac, ( C ) Doxorubicin + VLP-saLac, ( D ) Irinotecan, ( E ) Irinotecan + Lac, ( F ) Irinotecan + VLP-saLac; ( G ) Procarbazine, ( H ) Procarbazine + Lac, ( I ) Procarbazine + VLP-saLac. The percentage in every square indicates the percentage of total events.

Article Snippet: The MOLT-4 cell line, derived from human T cells originating from acute lymphoblastic leukemia, was obtained from a certified cell bank (ATCC, MOLT-4-CRL-1582).

Techniques: Flow Cytometry, Staining