rabbit polyclonal anti zo 2 antibody  (Santa Cruz Biotechnology)

Journal: Pathogens

Article Title: Role of Rab13, Protein Kinase A, and Zonula Occludens-1 in Hepatitis E Virus Entry and Cell-to-Cell Spread: Comparative Analysis of Quasi-Enveloped and Non-Enveloped Forms

doi: 10.3390/pathogens13121130

Figure Lengend Snippet: Impact of Rab13 silencing on HEV infection. ( A ) Schematic of the experimental protocol. PLC/PRF/5 cells were transfected with siRNA at two time points: 3 days prior to and 4 days post-HEV inoculation. HEV replication was monitored for up to 10 days post-inoculation (dpi). ( B , C ) Quantification of HEV RNA in the culture supernatants of cells inoculated with eHEV ( B ) or neHEV ( C ) and treated with siRab13. Data are presented as the mean ± standard deviation (SD) from three independent experiments. * p < 0.01. ( D , E ) Intracellular expression of HEV ORF2 and ORF3 proteins and knockdown efficiency of Rab13-specific siRNA (siRab13). PLC/PRF/5 cells were transfected with siRab13, non-targeting control siRNA (siNC), or buffer only (no siRNA). Cells were lysed on the indicated days following inoculation with eHEV ( D ) or neHEV ( E ), and the expression levels of ORF2 protein, ORF3 protein, Rab13, and β-actin were assessed by Western blotting using anti-ORF2 MAb, anti-ORF3 MAb, anti-Rab13 PAb, and anti-β-actin MAb, respectively.

Article Snippet: The resolved proteins were transferred to polyvinylidene difluoride (PVDF) membranes (0.45 μm; Merck Millipore) and probed with primary antibodies including anti-ORF2 mouse monoclonal antibody (MAb; H6225) [ ], anti-ORF3 mouse MAb (TA0536) [ ], anti-Rab13 rabbit polyclonal antibody (PAb; Merck Millipore), anti-ZO-1 rabbit PAb (Proteintech, Rosemont, IL, USA), anti-ZO-2 rabbit PAb (Cell Signaling Technology, Danvers, MA, USA), anti-ZO-3 rabbit MAb (Cell Signaling Technology), or anti-β-actin mouse MAb (Fujifilm Wako).

Techniques: Infection, Transfection, Standard Deviation, Expressing, Knockdown, Control, Western Blot

Journal: Pathogens

Article Title: Role of Rab13, Protein Kinase A, and Zonula Occludens-1 in Hepatitis E Virus Entry and Cell-to-Cell Spread: Comparative Analysis of Quasi-Enveloped and Non-Enveloped Forms

doi: 10.3390/pathogens13121130

Figure Lengend Snippet: The role of Rab13 in the HEV life cycle. ( A ) Schematic of the experimental design for the HEV-nanoKAZ assay. PLC/PRF/5 cells were transfected with siRNA 3 days prior to virus inoculation. Cells were then infected with eHEV-nanoKAZ or neHEV-nanoKAZ, and cell lysates were collected 4 dpi. ( B ) Intracellular luciferase activity in siRab13-treated cells inoculated with eHEV-nanoKAZ ( left panel ) or neHEV-nanoKAZ ( right panel ) at 4 dpi. Luciferase activity was measured using h-coelenterazine. ( C ) Schematic of the experimental design for the HEV-GLuc and HEV-HiBiT assays. PLC/PRF/5 cells were transfected with siRNA 3 days before transfection with pHEV3b-GLuc RNA or pHEV3b-HiBiT/ΔORF2s RNA. Following a 4 h inoculation at 37 °C, the culture medium was replaced with fresh growth medium, and the cells were incubated at 35.5 °C. Culture supernatants were collected 4 days post-transfection (dpt). ( D ) Luciferase activity in the culture supernatants of siRab13-treated cells transfected with pHEV3b-GLuc RNA ( left panel ) or pHEV3b-HiBiT/ΔORF2s RNA ( right panel ) at 4 dpt. Luciferase activity derived from GLuc was measured using coelenterazine, while HiBiT activity was assessed with the Nano-Glo HiBiT extracellular detection system after treatment of the culture supernatant with 1% digitonin. ( E ) Efficiency of Rab13 knockdown by siRNA. Cells were lysed 4 days after inoculation with eHEV or neHEV, or 4 days after transfection with pHEV3b-GLuc RNA or pHEV3b-HiBiT/ΔORF2s RNA. Expression levels of Rab13 ( upper panels ) and β-actin ( lower panels ) were analyzed by Western blotting using anti-Rab13 PAb and anti-β-actin MAb, respectively. All data are presented as the mean ± SD of three independent experiments. * p < 0.001.

Article Snippet: The resolved proteins were transferred to polyvinylidene difluoride (PVDF) membranes (0.45 μm; Merck Millipore) and probed with primary antibodies including anti-ORF2 mouse monoclonal antibody (MAb; H6225) [ ], anti-ORF3 mouse MAb (TA0536) [ ], anti-Rab13 rabbit polyclonal antibody (PAb; Merck Millipore), anti-ZO-1 rabbit PAb (Proteintech, Rosemont, IL, USA), anti-ZO-2 rabbit PAb (Cell Signaling Technology, Danvers, MA, USA), anti-ZO-3 rabbit MAb (Cell Signaling Technology), or anti-β-actin mouse MAb (Fujifilm Wako).

Techniques: Transfection, Virus, Infection, Luciferase, Activity Assay, Incubation, Derivative Assay, Knockdown, RNA Expression, Western Blot

Journal: bioRxiv

Article Title: A ZO-2 scaffolding mechanism regulates the Hippo signalling pathway

doi: 10.1101/2024.10.17.618965

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ZO-2 antibody (H-110), rabbit polyclonal anti-ZO-1 antibody (H-300), and rabbit polyclonal anti-Sp1 antibody (H-225) were purchased from Santa Cruz Biotechnology.

Techniques: shRNA