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