Journal: bioRxiv

Article Title: Sustained interferon exposure creates a hyper-metastatic subset of melanoma cells

doi: 10.64898/2026.04.01.715921

Figure Lengend Snippet: Higher interferon-regulated gene expression in metastasizing melanoma cells and increased formation of metastatic tumors after interferon treatment. a, We performed RNA sequencing on patient-derived xenograft cells (M405 and M481) isolated by flow cytometry from subcutaneous tumors, the blood, and metastatic tumors in NSG mice. After eliminating cell cycle-related genes, the 10 most significantly enriched gene sets in melanoma cells from the blood included ‘viral genome replication’ (red), which contains interferon-regulated genes. b, Interferon-regulated genes were more highly expressed by circulating (CMC) and metastatic as compared to primary subcutaneous (SQ) melanoma cells by gene set variation analysis (“Response to type I interferon” gene set). c-e, By qRT-PCR, transcript levels for the interferon-regulated genes ISG15, IFI27 and IFITM3 were higher in melanoma cells isolated from the blood as compared to subcutaneous or metastatic tumors of xenografted mice (two to three independent experiments per melanoma with a total of 3-5 mice per melanoma). f, Luciferase-expressing human melanoma cells were cultured overnight in 10 ng/mL hIFNa2 or vehicle and then intravenously injected into NSG mice. Metastatic disease burden was assessed five to nine weeks later by bioluminescence imaging of visceral organs and normalized to controls (two experiments per melanoma with a total of nine to ten mice per melanoma). g-j, Luciferase-expressing YUMM1.7, YUMM3.3, or YUMM5.2 mouse melanoma cells were cultured overnight in 10 ng/mL mIFN51, mIFNa2, mIFNy , or vehicle control and then injected subcutaneously (g, i) or intravenously (h, j) into NSG (g-h) or C57BL mice (i-j). We observed no differences in subcutaneous tumor growth (representative data are shown for YUMM1.7 cells; g, i). Interferon-treated mice gave rise to significantly higher tumor burden after intravenous injection into C57BL but not NSG mice (two independent experiments with a total of 9-20 mice per melanoma). k-n, IFNAR mutant or control mouse melanoma cells were injected subcutaneously (k, m) or intravenously (l, n) into NSG (k, l) or C57BL mice (m, n). IFNAR mutant and control cells did not differ in the size of the subcutaneous tumors they formed in NSG (k) or C57BL (m) mice (one experiment using YUMM1.7 cells is shown for each mouse strain, representative of 2 to 3 independent experiments for each of 3 melanomas). IFNAR mutant cells generally gave rise to fewer tumors in visceral organs than control cells after intravenous injection in NSG (l) and C57BL (n) mice, though the difference was much greater in C57BL mice. For each melanoma, two control clones and three independently-targeted IFNAR1 mutant clones were studied in two to four independent experiments per melanoma with a total of 10-25 mice per melanoma. Each dot represents a different mouse and all data represent mean ± s.d. Statistical significance was assessed using repeated measures one-way (c) or two-way ANOVAs (d-e) with Dunnett’s multiple comparisons adjustments (c-e), Mann-Whitney tests followed by Holm-Sidak’s multiple comparisons adjustments (f, Y3.3 and Y5.2 of h and j, l, and n), nparLD tests (g, i, k, and m) followed by FDR multiple comparisons adjustments (g and i), or Kruskal-Wallis tests with Dunn’s multiple comparisons adjustments (for Y1.7 of h and j). All statistical tests were two-sided. No statistically significant differences were observed in f, g-i, k, or m.

Article Snippet: YUMM 1.7 ( Braf V600E/+ ; PTEN -/- ; Cdkn2 -/- ), YUMM3.3 ( Braf V600E/+ ; Cdkn2 -/- ), and YUMM5.2 (Braf V600E/+ p53 -/- ) mouse melanomas were obtained from, and authenticated by, the American Type Culture Collection (ATCC).

Techniques: Gene Expression, RNA Sequencing, Derivative Assay, Isolation, Flow Cytometry, Quantitative RT-PCR, Luciferase, Expressing, Cell Culture, Injection, Imaging, Control, Mutagenesis, Clone Assay, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: SLy1-deficiency results in functional impaired, exhausted and senescent NK cells

doi: 10.3389/fimmu.2026.1836862

Figure Lengend Snippet: Phenotypic and functional effects of a SLy1 knockout on splenic NK cells in dependence on p53. (A) Absolute NK cell count determined via flow cytometry in the spleen of mice of the specified genotypes. (B) Apoptotic (defined as Annexin V + ) NK cells. (C) Living (defined as 7-AAD - ) NK cells after four hours under culture conditions. (D) Expression of NKG2D, NK1.1 and NKp46 on NK cells. (E) In vitro cytotoxicity against LLC at a 25:1 effector:target ratio. Dead LLC cells were defined as 7-AAD + /Annexin V + . In all figures error bars represent standard error of the mean. ∗: p <0.05, ∗∗/##: p <0.01, ns, not significant; unpaired t-test. (F) Freshly isolated NK cells (1) attack and lyse LLC cells (2) in vitro (see arrow 3). Image was taken after four hours of incubation. Scale bar = 20 µm.

Article Snippet: SLy1 KO mice were further crossed with a floxed p53 mouse strain (B6.129P2- Trp53 tm1Brn /J; strain #008462 Jackson Laboratory).

Techniques: Functional Assay, Knock-Out, Cell Characterization, Flow Cytometry, Expressing, In Vitro, Isolation, Incubation

Journal: Frontiers in Immunology

Article Title: SLy1-deficiency results in functional impaired, exhausted and senescent NK cells

doi: 10.3389/fimmu.2026.1836862

Figure Lengend Snippet: Senescent (C 12 FDG + ) NK cells in the spleen (upper part) and lungs (lower part). (A, C) Comparison of the proportion of senescent NK cells in young (8 weeks) and old (20 weeks) mice depending on SLy1 genotype. (B, D) Senescence (in 20-week-old mice) in SLy1 knockout NK cells depending on p53. Error bars represent standard error of the mean. ∗/#, p <0.05; ∗∗/##, p <0.01; ns, not significant; unpaired t-test.

Article Snippet: SLy1 KO mice were further crossed with a floxed p53 mouse strain (B6.129P2- Trp53 tm1Brn /J; strain #008462 Jackson Laboratory).

Techniques: Comparison, Knock-Out

Journal: Frontiers in Immunology

Article Title: SLy1-deficiency results in functional impaired, exhausted and senescent NK cells

doi: 10.3389/fimmu.2026.1836862

Figure Lengend Snippet: Schematic presentation of the effect of SLy1-deficiency on the phenotype and function of NK cells and the relationship to p53. Increase in free ribosomal proteins was previously shown by Arefanian et al. Dashed lines represent hypotheses. Created in BioRender . Beer-Hammer, S. ( https://BioRender.com/hjgbedc ) is licensed under CC BY 4.0.

Article Snippet: SLy1 KO mice were further crossed with a floxed p53 mouse strain (B6.129P2- Trp53 tm1Brn /J; strain #008462 Jackson Laboratory).

Techniques:

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and p53 after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.

Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

Techniques: Western Blot, Transfection, Expressing, Control

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.

Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

Techniques: Western Blot, Inhibition, Pull Down Assay, Incubation, Purification, Staining

Journal: eBioMedicine

Article Title: Modified hTERT treatment ameliorates pressure overload-induced heart failure

doi: 10.1016/j.ebiom.2026.106203

Figure Lengend Snippet: JV101 inhibited telomeric DNA damage, mitochondrial dysfunction and chronic inflammation in TAC-induced HF. (A) Chord diagrams showing differentially expressed genes corresponding to selected regulated pathways in TAC + JV101 versus TAC + Vector comparisons. (B and C) Immunoblotting analysis of multiple inflammatory cytokines in heart of TAC-induced heart failure mice (n = 3 per group). (D) Nuclear lysate isolated from cardiomyocytes perfused from mice of TAC + Vector, TAC + JV101 and Sham group were immunoblotted for gH2AX, Histone H2AX, and 53BP1; Whole cell lysate was immunoblotted for p-p53(Ser15), p53, PGC1 a and TFAM. H3 and GAPDH were used as loading controls. (E–G) Representative transmission electron micrographs of left ventricular myocardium of the indicated study groups. Quantitative analyses of cristae number (F) and mitochondria density (G) in TAC hearts. (H) Quantification of mitochondrial DNA (mtDNA) copy numbers of cardiomyocytes perfused from mice of TAC + Vector, TAC + JV101 and Sham group (n = 6). (I) Intracellular ATP content of TAC + Vector, TAC + JV101 and Sham group (n = 6). Data are represented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Statistical differences among groups were analysed by one-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: The primary antibodies: histone H 2 AX (1:1000, Cell Signalling Technology, 7631S), γH 2 AX (1:1000, Cell Signalling Technology, 9718S), 53BP1(1:1000, Cell Signalling Technology, 4937S), p53 (1:1000, Cell Signalling Technology, 2524S), Phospho-p53 (Ser15) (1:1000, Cell Signalling Technology, 9284s), PGC-1α (1:1000, Santa Cruz, sc-517380), TFAM (1:1000, Proteintech, 22586-1-AP), TRF1 (1:1000, Abcam, ab192629), RAP1 (1:1000, Santa Cruz, sc-53434), TIN2 (1:1000, Proteintech, 11368-1-AP), POT1 (1:1000, Novus Biologicals, NB500-176), HA-tag (1:1000, Cell Signalling Technology, 3724s), DYKDDDDK Tag (1:1000, Cell Signalling Technology, 14793s), H3 (1:1000, Cell Signalling Technology, D5A7), GAPDH (1:1000, Proteintech, 60004-1-Ig) followed by secondary antibody for 1 h with a 1:2000 dilution of IgG Goat Anti-Mouse HRP (Proteintech, SA00001-1) or IgG Goat Anti- Rabbit HRP (Proteintech, SA00001-2).

Techniques: Plasmid Preparation, Western Blot, Isolation, Transmission Assay

Journal: eBioMedicine

Article Title: Modified hTERT treatment ameliorates pressure overload-induced heart failure

doi: 10.1016/j.ebiom.2026.106203

Figure Lengend Snippet: JV101 blocked DDR-p53-TFAM axis and inflammation response in Ang II-induced hiPSC-CMs. (A) Expression levels of inflammatory cytokines in hiPSC-CMs were determined by RT-qPCR (n = 6). (B) Representative micrographs of hiPSC-CMs displaying Telomere (red), 53BP1 (green) and DAPI (blue). (C) Quantification of 53BP1-telomere co-localisation (n = 6). (D) Cell lysate isolated from Ang II + Vector, Ang II + JV101 and control hiPSC-CMs were immunoblotted for p-p53(Ser15), p53, PGC-1a and TFAM. (E–G) Representative transmission electron micrographs of left ventricular myocardium of the indicated study groups. Quantitative analyses of cristae number (F) and mitochondria density (G) in hiPSC-CMs. (H) Quantification of mitochondrial DNA (mtDNA) copy numbers in different groups (n = 6). (I) Intracellular ATP content of different groups (n = 6). Data are represented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Statistical differences among groups were analysed by one-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: The primary antibodies: histone H 2 AX (1:1000, Cell Signalling Technology, 7631S), γH 2 AX (1:1000, Cell Signalling Technology, 9718S), 53BP1(1:1000, Cell Signalling Technology, 4937S), p53 (1:1000, Cell Signalling Technology, 2524S), Phospho-p53 (Ser15) (1:1000, Cell Signalling Technology, 9284s), PGC-1α (1:1000, Santa Cruz, sc-517380), TFAM (1:1000, Proteintech, 22586-1-AP), TRF1 (1:1000, Abcam, ab192629), RAP1 (1:1000, Santa Cruz, sc-53434), TIN2 (1:1000, Proteintech, 11368-1-AP), POT1 (1:1000, Novus Biologicals, NB500-176), HA-tag (1:1000, Cell Signalling Technology, 3724s), DYKDDDDK Tag (1:1000, Cell Signalling Technology, 14793s), H3 (1:1000, Cell Signalling Technology, D5A7), GAPDH (1:1000, Proteintech, 60004-1-Ig) followed by secondary antibody for 1 h with a 1:2000 dilution of IgG Goat Anti-Mouse HRP (Proteintech, SA00001-1) or IgG Goat Anti- Rabbit HRP (Proteintech, SA00001-2).

Techniques: Expressing, Quantitative RT-PCR, Isolation, Plasmid Preparation, Control, Transmission Assay