Western Blot Analysis of Cell Lysates

Cells were harvested and lysed and protein content of cell lysates was measured as described previously [26 (link)] Whole cell lysates were separated by SDS-PAGE (in 10% or 15% SDS-polyacrilamide gel). SDS-PAGE and Western Blot analysis was done according to our previous study [26 (link)]The following antibodies were applied: anti-NQO1 (Cell Signaling, A180), anti-CyclinD1 (Santa Cruz, A-12), anti-p21 (Santa Cruz, C-19), anti-p27 (Santa Cruz, C-19), anti-p15-INK4b (proteintech, 12877-1-AP), anti-p16-INK4A (proteintech, 10883-1-AP), anti-PERK (Cell Signaling, 3192S), anti-P-eIF2α (Cell Signaling, 9722S9), anti-P-eIF2α (Ser51) (Cell Signaling, 9721L), and anti-GAPDH (Santa Cruz, 6C5) and HRP conjugated secondary antibodies (SantaCruz, sc-2020 and Cell Signaling, 7074S, 7076S).

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Márton M., Tihanyi N., Gyulavári P., Bánhegyi G, & Kapuy O. (2018). NRF2-regulated cell cycle arrest at early stage of oxidative stress response mechanism. PLoS ONE, 13(11), e0207949.

Publication 2018

anti-NQO1 anti-CyclinD1 anti-p21 anti-p27 anti-p16-INK4A anti-PERK anti-P-eIF2α anti-P-eIF2α (Ser51) anti-GAPDH

Antibodies Cell Gapdh Nqo1 P16 ink4a Protein Sds page Western blot analysis

Corresponding Organization : Semmelweis University

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Variable analysis

independent variables

Cells were harvested and lysed

dependent variables

Protein content of cell lysates
Expression levels of NQO1, CyclinD1, p21, p27, p15-INK4b, p16-INK4A, PERK, P-eIF2α, and GAPDH proteins

control variables

Experimental procedures were done according to a previous study [26]

controls

Positive control: GAPDH (housekeeping gene)
Negative control: Not explicitly mentioned

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