Journal: bioRxiv

Article Title: HPV Capsid-Derived Cationic Peptides for Cargo Delivery and Antiviral Activity

doi: 10.64898/2026.05.06.723171

Figure Lengend Snippet: | WT peptide internalization is dependent on the overall peptide sequence and the positively charged residues. HaCaT cells were treated with 4.5 μM of the indicated peptide for 2 h and analyzed by flow cytometry. (A) Representative histograms show the percentage of FITC-positive HaCaT cells after treatment with FITC-SC and FITC-WT peptide. (B) Quantification of the average percentage. (C) Representative flow cytometry histogram showing the percentage of FITC-positive HaCaT cells after treatment with M1, M2, and FITC-WT peptides. (D) Quantification of the average percentage. Data are represented as mean ± SEM (n=3). Data were analyzed by unpaired two-tailed Student’s t -test; *, P <0.05; **, P <0.01; ***, P <0.001; ns , p>0.05.

Article Snippet: The next day the cell were treated with pre-treated with 20 μM chlorpromazine (Sigma Aldrich; St. Louis, MO, USA; C8313), 2 mM methyl-β-cyclodextrin (MβCD) (Sigma Aldrich; St. Louis, MO, USA; C4555), or 50 μM Dynasore (Sigma Aldrich; St. Louis, MO, USA; D7693) for 2 h, following the pre-incubation the cells were treated with 4.5 μM FITC-WT peptide, Human Transferrin CF488A (Biotium, 00081), or cholera toxin subunit B conjugated to Alexa Fluor-488 (Invitrogen, Thermo Fisher Scientific, C34775) in the presence of inhibitors for an additional 2 h. The cells were then trypsinized and subjected to flow cytometry.

Techniques: Sequencing, Flow Cytometry, Two Tailed Test

Journal: bioRxiv

Article Title: HPV Capsid-Derived Cationic Peptides for Cargo Delivery and Antiviral Activity

doi: 10.64898/2026.05.06.723171

Figure Lengend Snippet: | FITC-WT peptide internalization is mediated by heparan sulfate via energy-dependent endocytosis. (A) HaCaT cells treated with Buffer (control) or Heparinase for 2 h at 37 °C. The cells were then treated with 4.5 μM FITC-WT for 2 h at 37 °C. Internalization was analyzed using flow cytometry. Representative histograms show the percentage of FITC-positive cells under each condition. (B) Quantification of the average percentage of FITC-positive HaCaT cells. (C) HaCaT cells were treated with 4.5 μM FITC-WT for 2 h at 4 °C or 37 °C, followed by analysis via flow cytometry. Representative histograms show the percentage of FITC-positive cells. (D) Quantification of the average percentage of FITC-positive HaCaT cells at 4 °C and 37 °C after 2 h. (E) Confocal fluorescence microscopy images of HaCaT cells treated with 4.5 μM FITC or FITC-WT for 2 h at 37 °C. Gray corresponds to DAPI (nuclear stain), green corresponds to FITC-WT, and red corresponds to EEA1 (early endosomal marker). Scale bar: 20 μm. Data are represented as mean ± SEM (n=3). Data were analyzed by unpaired two-tailed Student’s t -test; *, P <0.05; **, P <0.01; ***, P <0.001; ns , p>0.05.

Article Snippet: The next day the cell were treated with pre-treated with 20 μM chlorpromazine (Sigma Aldrich; St. Louis, MO, USA; C8313), 2 mM methyl-β-cyclodextrin (MβCD) (Sigma Aldrich; St. Louis, MO, USA; C4555), or 50 μM Dynasore (Sigma Aldrich; St. Louis, MO, USA; D7693) for 2 h, following the pre-incubation the cells were treated with 4.5 μM FITC-WT peptide, Human Transferrin CF488A (Biotium, 00081), or cholera toxin subunit B conjugated to Alexa Fluor-488 (Invitrogen, Thermo Fisher Scientific, C34775) in the presence of inhibitors for an additional 2 h. The cells were then trypsinized and subjected to flow cytometry.

Techniques: Control, Flow Cytometry, Fluorescence, Microscopy, Staining, Marker, Two Tailed Test

Journal: bioRxiv

Article Title: HPV Capsid-Derived Cationic Peptides for Cargo Delivery and Antiviral Activity

doi: 10.64898/2026.05.06.723171

Figure Lengend Snippet: | FITC-WT peptide internalizes via lipid-raft endocytosis, independent of Clathrin, Caveolae, and Dynamin. (A) HaCaT cells were either pretreated with vehicle, DMSO, or 20 μM chlorpromazine, followed by treatment with either FITC-WT peptide or transferrin, positive control, for 2 h at 37 °C in the presence of the inhibitor. Internalization was analyzed using flow cytometry. Representative histograms show the percentage of FITC-positive cells. (B) Quantification of the average percentage of FITC-positive cells. (C) HaCaT cells were either pretreated with vehicle, H 2 O, or 2 mM MβCD, followed by treatment with either FITC-WT peptide or CTB, positive control, for 2 h at 37 °C in the presence of the inhibitor. Internalization was analyzed via flow cytometry. Representative histograms show the percentage of FITC-positive cells. (D) Quantification of the average percentage of FITC-positive cells. (E) HaCaT cells were either pretreated with vehicle, DMSO, or 50 μM dynasore, followed by treatment with either FITC-WT peptide or transferrin, positive control, for 2 h at 37 °C in the presence of the inhibitor. Internalization was analyzed via flow cytometry. Representative histograms show the percentage of FITC-positive cells. (F) Quantification of the average percentage of FITC-positive cells. Data are represented as mean ± SEM (n=3). Data were analyzed by unpaired two-tailed Student’s t -test; *, P <0.05; **, P <0.01; ***, P <0.001; ns , p>0.05.

Article Snippet: The next day the cell were treated with pre-treated with 20 μM chlorpromazine (Sigma Aldrich; St. Louis, MO, USA; C8313), 2 mM methyl-β-cyclodextrin (MβCD) (Sigma Aldrich; St. Louis, MO, USA; C4555), or 50 μM Dynasore (Sigma Aldrich; St. Louis, MO, USA; D7693) for 2 h, following the pre-incubation the cells were treated with 4.5 μM FITC-WT peptide, Human Transferrin CF488A (Biotium, 00081), or cholera toxin subunit B conjugated to Alexa Fluor-488 (Invitrogen, Thermo Fisher Scientific, C34775) in the presence of inhibitors for an additional 2 h. The cells were then trypsinized and subjected to flow cytometry.

Techniques: Positive Control, Flow Cytometry, Two Tailed Test

Journal: Blood

Article Title: NLRP3 inflammasome blockade treats intestinal inflammation associated with chronic granulomatous disease

doi: 10.1182/blood.2025029676

Figure Lengend Snippet: βHB inhibits NLRP3 inflammasome activity in human cells in the context of CGD. Western blot analysis of IL-1β (pro–IL-1β and active IL-1β) and β-actin (loading control) expression in WT (left) compared to CYBB −/− (ie, CGD; right) THP-1 cells (A) or PBMCs from a healthy person and a patient with CGD (C) that were either unstimulated, treated with the nigericin trigger (1 hour) alone (nigericin), or stimulated with lipopolysaccharide (LPS; 3 hours) and nigericin (1 hour) with or without NLRP3 inhibition with different doses of βHB (1mM-20mM) 30 minutes before adding the nigericin trigger. The last lane shows cells treated with 10mM βHB alone (ie, no priming and no trigger). The measurement of IL-1β, IL-6, and TNF levels in cell culture supernatants after the treatment of WT and CYBB −/− THP-1 cells (B) or healthy and CGD PBMCs (D) with either PBS (unstimulated), LPS (3 hours) and nigericin (1 hour; stimulated), or LPS followed by NLRP3 inhibition with 20mM βHB before (stimulated + βHB) or 10μM MCC950 (stimulated + MCC950) for 30 minutes before adding nigericin. Data in panels C-D are representative of 2 independent experiments. Significance was determined using multiple Mann-Whitney U tests for panel C or unpaired t tests for panel D, with data represented as median ± IQR or mean ± SEM, respectively. Stim., stimulated; Unstim., unstimulated.

Article Snippet: In vitro experiments were performed using human monocytic leukemia THP-1 cell lines, including WT THP-1 (TIB-202; American Type Culture Collection) and NOX2-deficient ( CYBB −/− ) THP-1 cells (generously provided by Fabien Touzot, CHU Sainte-Justine ), cultured in RPMI 1640 (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Gibco) and 1% penicillin-streptomycin-glutamine (Gibco).

Techniques: Activity Assay, Western Blot, Control, Expressing, Inhibition, Cell Culture, MANN-WHITNEY

Journal: HemaSphere

Article Title: Casein kinase 1δ/ε inhibition suppresses CLL proliferation through cell‐intrinsic and microenvironmental mechanisms

doi: 10.1002/hem3.70343

Figure Lengend Snippet: Validation of the cell cycle and proliferation effects of casein kinase 1δ/ε (CK1δ/ε) inhibition in vivo and in vitro. (A) Percentages of EdU‐Alexa Fluor 647+ leukemic B cells within the spleen (SPL) of treated and control TCL1 adoptive transfer (AT) recipient mice ( N (AT CTRL) = 3; N (AT + PF‐670462) = 4), tested by the t ‐test. (B) Relative cell counts (% of CTRL) originating from in vitro treated MEC‐1 wild‐type (WT) cells after a 72 h treatment with PF‐670462 or MU1742 (performed on the following biological replicates: N (CTRL) = 6; N (3µM PF‐670462) = 3; N (10µM PF‐670462) = 6; N (3µM MU1742) = 3; and N (10µM MU1742) = 3), tested by the Kruskal–Wallis test with post hoc pairwise Wilcoxon rank sum tests with Benjamini–Hochberg correction. (C) Relative cell counts (% of CTRL) originating from in vitro treated HG‐3 WT cells after 72 h treatment with PF‐670462 or MU1742 (performed on the following biological replicates: N (CTRL) = 4; N (3µM PF‐670462) = 4; N (10µM PF‐670462) = 4; N (3µM MU1742) = 4; and N (10µM MU1742) = 4), tested by the Kruskal–Wallis test with post hoc pairwise Wilcoxon rank sum tests with Benjamini–Hochberg correction. (D) Cell cycle assay setup with initial CK1 inhibitor treatment and mitotic arrest with nocodazole and the representative example of cell cycle alterations between analyzed conditions in MEC‐1 and HG‐3 cell lines. (E) Cell cycle phase distribution in MEC‐1 WT cells upon 9 h pre‐treatment with 3 µM PF‐670462 and 10 µM PF‐670462 and subsequent mitotic arrest with nocodazole (performed on the following biological replicates: N (CTRL) = 11; N (NOCODAZOLE) = 11; N (3µM PF‐670462) = 7; and N (10µM PF‐670462) = 11); for all cases together, the generalized linear mixed‐effects model, followed by estimated marginal means calculation (P‐value < 0.05), was used separately for comparison of CTRL versus NOCODAZOLE and NOCODAZOLE versus PF‐670462 and corrected due to usage of 2 models. (F, G) Cell cycle phase distribution in MEC‐1 WT cells upon 9 h pre‐treatment with 3 and 10 µM concentrations of MU1742 and AH078 (respectively) and subsequent mitotic arrest with nocodazole (performed on the following biological replicates: N (CTRL) = 11; N (NOCODAZOLE) = 11; N (3µM) = 3; and N (10µM) = 3); for all cases together, the generalized linear mixed‐effects model followed by estimated marginal means calculation (P‐value < 0.05), was used separately for comparison of CTRL versus NOCODAZOLE and NOCODAZOLE versus MU1742/AH078 and corrected due to usage of 2 models. (H–J) Cell cycle phase distribution in HG‐3 WT cells upon 9 h pre‐treatment with 3 and 10 µM concentrations of PF‐670462, MU1742, and AH078 (respectively) and subsequent mitotic arrest with nocodazole (performed on the following biological replicates: N (CTRL) = 4; N (NOCODAZOLE) = 4; N (3µM) = 4; and N (10µM) = 4); for all cases together, the generalized linear mixed‐effects model, followed by estimated marginal means calculation (P‐value < 0.05), was used separately for comparison of CTRL versus NOCODAZOLE and NOCODAZOLE versus PF‐670462/MU1742/AH078 and corrected due to usage of 2 models. DMSO, dimethyl sulfoxide; PI, propidium iodide.

Article Snippet: CLL cell lines MEC‐1 WT (DSMZ, #ACC497) and HG‐3 WT (DSMZ, #ACC765) were treated with PF‐670462 (DC Chemicals, #DC2086), an in‐house CK1δ/ε inhibitor MU1742 or CK1δ/ε degrader AH078, and subjected to cell proliferation tracking and cell cycle tracking via PI staining, EdU Click‐iT assays, and/or western blotting, as described in more detail in the Supporting Information S1: .

Techniques: Biomarker Discovery, Inhibition, In Vivo, In Vitro, Control, Adoptive Transfer Assay, Cell Cycle Assay, Comparison