4.7.4. Western Blot Analysis

Protein expression levels of catalase (CAT), superoxide dismutase (SOD), nuclear factor E2-related factor 2 (Nrf2), and Kelch-like ECH-associated protein 1 (Keap1) were analyzed via western blotting. Cells were seeded in 6 well culture plates and samples were treated given 24 h period, following 1 h incubation cells were stimulated with H2O2. Cells were harvested after complete incubation and lysed. Protein content was measured and standardized. Following electrophoresis, it was transferred on to nitrocellulose membranes. Blocked membranes (5% skim milk) were incubated with primary and secondary antibodies step wisely (Santa Cruz Biotechnology, Paso Robles, CA, USA). The bands were developed and photographed via a FUSION SOLO Vilber Lourmat system. ImageJ program was assisted in the quantification of the band intensities [55,56].

#### *4.8. In Vivo Antioxidant E*ff*ects of PBP Using Zebrafish Model*
