Journal: Oncology Letters

Article Title: A genome-wide Mendelian randomization study focusing on endoplasmic reticulum stress reveals novel genetic markers for renal cell carcinoma

doi: 10.3892/ol.2026.15732

Figure Lengend Snippet: ER stress induces RCC development and resistance to sunitinib. (A) qPCR (left) and TEM (right) confirming ER stress induction in RCC development. In total, 3 cell line models were used: HK-2 (normal renal epithelial cells), 293T with adriamycin (nephropathy model) and 786-O (clear cell RCC). qPCR markers for ER stress included GRP78, PERK, IRE1α, ATF6 and CHOP. TEM scale bar, 500 nm scale. The levels of ER stress markers are higher in sunitinib resistant (B) 786-O and (C) Caki-1 cells. ER stress phenotype was verified by qPCR (left) and TEM (right, 500 nm scale). (D) Western blot analysis of ER stress markers (p-PERK, PERK, p-IRE1α, IRE1α, p-eIF2α, eIF2α, GRP78, CHOP and ATF6) in 786-O and Caki-1 cell lines with or without sunitinib resistance, showing activation of ER stress pathways. ACTB served as the loading control. Treatment with 4-PBA (ER stress inhibitor) was included as an experimental control. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. ER, endoplasmic reticulum; TEM, transmission electron microscopy; RCC, renal cell carcinoma; qPCR, quantitative PCR; GRP78, glucose-regulated protein 78; PERK, protein kinase R-like endoplasmic reticulum kinase; IRE1α, inositol-requiring enzyme 1α; ATF6, activating transcription factor 6; CHOP, C/EBP homologous protein; ACTB, actin beta; p-, phosphorylated.

Article Snippet: For ER stress inhibition, cells were pretreated with 4-PBA (2 mM, 24 h, at 37°C in a 5% CO 2 atmosphere; MedChemExpress; cat. no. HY-A2081) prior to protein extraction.

Techniques: Western Blot, Activation Assay, Control, Transmission Assay, Electron Microscopy, Real-time Polymerase Chain Reaction

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