Journal: bioRxiv

Article Title: Peroxisome dynamics during HSV-1 life cycle in human neurons

doi: 10.64898/2026.06.23.732381

Figure Lengend Snippet: a. Brief description of the workflow. SH-SY5Y cells were pretreated or not with 4-PBA for 48 hours and with NCC 55-0936 for 24 hours. Then, the cells were infected with HSV-1 strain McIntyre at an MOI of 0.1 and analyzed using high content confocal microscopy, viral yield and western blot analysis. b. Panel of confocal microscopy images representing the diQerent conditions. Peroxisomes were labelled using PEX14 (orange), HSV-1 using HSV-1 gD (green), nuclei using DAPI (blue), and cytoskeleton using phalloidin (red). Images were acquired using a 40x magnification. Each data point represents the mean value obtained analysing > 8×10 3 cells. c. Analysis of number of peroxisomes per cell after the treatment with 4-PBA and DMSO. d. Morphological analysis of peroxisomes expressed as spot area, width to length ratio, and roundness after the treatment with 4-PBA. e. Analysis of the percentage of infected cells 4-PBA and NCC 55-0396 treated compared to the vehicle treated control. f. Analyses of peroxisomes per cell after the treatment with NCC 55-0396 and DMSO as vehicle control. g. Viral titer of the cell treated with 4-PBA and DMSO expressed as PFU/mL. h. Viral titer of the cell treated with NCC 55-0396 and DMSO expressed as PFU/mL. i. Western blot analysis on HSV-1 gD content following treatments. GAPDH was used as loading control. Results are expressed as mean ± SD of independent replicates. Data were analysed with One-Way ANOVA (* p < 0.05, ** p < 0.01, *** p < 0.001; **** p < 0.0001)

Article Snippet: For peroxisome modulation, cells were treated with 3 mM 4-phenylbutiryc acid (4-PBA) (MedChemExpress, USA, #Cat 1821-12-1) for 48 hours or NCC 55-0396 (MedChemExpress, USA, #Cat 357400) for 24 hours before HSV-1 inoculation and after the infection.

Techniques: Infection, Confocal Microscopy, Western Blot, Control

Journal: bioRxiv

Article Title: Peroxisome dynamics during HSV-1 life cycle in human neurons

doi: 10.64898/2026.06.23.732381

Figure Lengend Snippet: a. Schematic representation of AGPS KO cell generation. b. Western blot image showing the AGPS KO generation. c. Schematic representation of the workflow. SH-SY5Y cells were pretreated or not with 4-PBA for 48 hours and with NCC 55-0936 for 24 hours. Then, the cells were infected with HSV-1 strain McIntyre at an MOI of 0.1 and analyzed using high content confocal microscopy and viral yield d. Panel of confocal microscopy images representing the diQerent conditions. HSV-1 using HSV-1 gD (red) and nuclei using DAPI (blue). Images were acquired using a 20x magnification. Each data point represents the mean value obtained analysing > 8×10 3 cells. e. Analysis of the % of infected cells among the diQerent conditions: SH-SY5Y WT, AGPS KO treated or not with 4-PBA. f. Representative images of viral yield assay plate for each analyzed condition. g. Quantification of HSV-1 viral titer expressed as PFU/mL in the diQerent samples. Each data point represents a single experiment.

Article Snippet: For peroxisome modulation, cells were treated with 3 mM 4-phenylbutiryc acid (4-PBA) (MedChemExpress, USA, #Cat 1821-12-1) for 48 hours or NCC 55-0396 (MedChemExpress, USA, #Cat 357400) for 24 hours before HSV-1 inoculation and after the infection.

Techniques: Western Blot, Infection, Confocal Microscopy