Western Blot Analysis of Paw and Spinal Cord

Western blot examination of the paw and spinal cord tissues was prepared as previously described in [72 (link)]. The following primary antibodies were used (for the purpose of standardization): anti-NF-κB (1:500), anti-Nrf-2 (1:500), anti-HO-1 (1:500), anti-IL-1β (1:500), anti-c-Fos (1:500), anti-βactin (1:500), and β-laminin (1:500) (Santa Cruz Biotechnology, Heidelberg, Germany) [73 (link)]. Protein expression was quantified via densitometry with Bio-Rad ChemiDocTM XRS+ software (Bio-Rad, Milan, Italy) and normalized with the housekeeping genes β-actin and lamin A/C, as previously reported in [74 (link),75 (link)].

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Interdonato L., Ferrario G., Cordaro M., D’Amico R., Siracusa R., Fusco R., Impellizzeri D., Cuzzocrea S., Aldini G, & Di Paola R. (2023). Targeting Nrf2 and NF-κB Signaling Pathways in Inflammatory Pain: The Role of Polyphenols from Thinned Apples. Molecules, 28(14), 5376.

Publication 2023

anti-NF-κB anti-Nrf-2 anti-HO-1 anti-IL-1β anti-c-Fos anti-βactin β-laminin Bio-Rad ChemiDocTM XRS+ software β-actin

Actin Antibodies Densitometry Housekeeping genes Il 1β Lamin a c Laminin Nf κb Nrf 2 Protein Spinal cord Tissues Western blot

Corresponding Organization : University of Messina

Other organizations : University of Milan

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Variable analysis

independent variables

Primary antibodies used: anti-NF-κB, anti-Nrf-2, anti-HO-1, anti-IL-1β, anti-c-Fos, anti-βactin, and β-laminin

dependent variables

Protein expression

control variables

Paw and spinal cord tissues prepared as previously described in [72]
Protein expression quantified via densitometry and normalized with the housekeeping genes β-actin and lamin A/C, as previously reported in [74,75]

controls

Positive control: None specified
Negative control: None specified

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