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: Journal of Nanobiotechnology

Article Title: Targeting the HDAC4-NHE6-endosomal pH axis restores amyloid-β clearance and cognitive function in Alzheimer's disease mice

doi: 10.1186/s12951-026-04297-2

Figure Lengend Snippet: SAHA@LIPO-ANG2 corrects endosomal pH, facilitating the release and subsequent cell membrane expression of LRP1. ( a – d ) Fluorescence images ( a ) and corresponding intensities ( c ) (n = 6) of BV2 (left) and C8-D1A (right) treated with different concentrations of LV-NHE6 KD-mCherry; flow cytometry results ( b ) (n = 3); comparison of transfection efficiencies between LV-NHE6 KD-mCherry and LV-mCherry groups ( d ). Scale bar, 200 μm. ( e , f ) Representative Western blotting of NHE6 and LRP1. Cells were collected after lentiviral treatment, lysed in RIPA buffer, and analyzed for protein expression in the supernatant ( e ) and quantitative analysis of relevant proteins ( f ). (n=4) ( g - i ) Fluorescence images of NHE6 knockdown and control group in BV2 and C8-D1A cells ( g ) (n = 6), cell (mCherry, red), DAPI (blue), and PH (FITC, green), flow cytometry results ( h ) (n=3), and corresponding intensities of fluorescence ( i ). C8-D1A represents astrocytes, BV2 represents microglia. ( j , k ) LRP1 (green) fluorescence images and corresponding intensities ( k ) in NHE6 knockdown and SAHA@LIPO-ANG2 treated groups (n=6). Statistical comparisons in ( c ) were performed using one-way ANOVA and two-way ANOVA, respectively. Statistical comparisons in ( d ), ( f ), ( i ), and ( k ) were performed using unpaired two-tailed t-test with Welch’s correction. (ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)

Article Snippet: The antibodies used are as follows: IBA1 (Oasis, OB-PGP049-01), Aβ (BioLegend, 800708), LRP1 (ZRB1311), GFAP (FujiFilm, MO389), β-amyloid-FITC (MCE, HY-P3908), LAMP2 (Proteintech, 66301-1-Ig), S100A10 (Proteintech, 11250-1-AP).

Techniques: Membrane, Expressing, Fluorescence, Flow Cytometry, Comparison, Transfection, Western Blot, Knockdown, Control, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Targeting the HDAC4-NHE6-endosomal pH axis restores amyloid-β clearance and cognitive function in Alzheimer's disease mice

doi: 10.1186/s12951-026-04297-2

Figure Lengend Snippet: SAHA@LIPO-ANG2 enhances microglial phagocytic activity. ( a ) Representative immunofluorescence images showing the co-localization of Aβ plaques (purple) and microglia (Iba1, green) in the cortex of 5xFAD mice treated with SAHA@LIPO-ANG2 or control. ( b ) Quantitative analysis of the co-localization between Aβ plaques and microglia. Increasing in Pearson’s correlation coefficient indicates enhanced co-localization. ( c ) Representative images of Aβ (purple), Iba1 (microglia, green), and CD68⁺ (red) in the cortical regions of the treatment and control groups; ( d ) linear fluorescence intensity profiles along the white dashed lines are shown on the left. ( e ) Confocal images of BV2 microglia co-cultured with FITC-Aβ42 for 4 hours following treatment with SAHA@LIPO-ANG2. ( f ) Quantification of FITC-Aβ42 internalization by BV2 microglia, analyzed using ImageJ software (n = 7–10 per group). A.U., arbitrary units. ( g ) Representative 3D-rendered image of Iba1 (red), CD68⁺ (green), and Aβ (cyan) by Imaris software. ( h ) Volume analysis of Aβ and microglia co-localization. ( i – m ) Representative Western blotting bands for CD36, APP, Aβ and GAPDH expression ( i ). Cortical and hippocampal tissues were collected after behavioral tests and lysed in RIPA buffer; proteins from the supernatant were quantified for APP( j ), Aβ( k ), CD36( l ) and GAPDH ( m ) (n = 4). Statistical comparisons for ( f ) and ( h ), data are presented as mean ± SEM, by Student’s t-test. ( j ), ( k ), ( l ) and ( m )were performed using one-way ANOVA (ns, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)

Article Snippet: The antibodies used are as follows: IBA1 (Oasis, OB-PGP049-01), Aβ (BioLegend, 800708), LRP1 (ZRB1311), GFAP (FujiFilm, MO389), β-amyloid-FITC (MCE, HY-P3908), LAMP2 (Proteintech, 66301-1-Ig), S100A10 (Proteintech, 11250-1-AP).

Techniques: Activity Assay, Immunofluorescence, Control, Fluorescence, Cell Culture, Software, Western Blot, Expressing