Cells were grown on
fibronectin-coated coverslips in the presence or absence of 6S. After
treatment with test samples for the indicated times, cells were fixed
with cold 2% (w/v) paraformaldehyde for 20 min, permeabilized in 0.1%
(w/v) Triton X-100 in 1× PBS, washed, and blocked in 1% BSA at
room temperature for 1 h. Tissue sample sections were deparaffinized
and rehydrated. After being washed with PBS for 5 min three times,
the cells were incubated with a rabbit anti-Nrf2 antibody (1:200;
Santa Cruz Biotechnology) and tissue sections were incubated with
a rabbit anti-Nrf2 antibody (1:200)39 (link) overnight
at 4 °C, followed by FITC-conjugated secondary antibody (1:200)
for 1 h at room temperature. Samples were counterstained with DAPI
(1 mg/mL) for visualization of the nuclei. Stained samples were mounted
and visualized under a fluorescent microscope (Thermo Fisher Scientific).
fibronectin-coated coverslips in the presence or absence of 6S. After
treatment with test samples for the indicated times, cells were fixed
with cold 2% (w/v) paraformaldehyde for 20 min, permeabilized in 0.1%
(w/v) Triton X-100 in 1× PBS, washed, and blocked in 1% BSA at
room temperature for 1 h. Tissue sample sections were deparaffinized
and rehydrated. After being washed with PBS for 5 min three times,
the cells were incubated with a rabbit anti-Nrf2 antibody (1:200;
Santa Cruz Biotechnology) and tissue sections were incubated with
a rabbit anti-Nrf2 antibody (1:200)39 (link) overnight
at 4 °C, followed by FITC-conjugated secondary antibody (1:200)
for 1 h at room temperature. Samples were counterstained with DAPI
(1 mg/mL) for visualization of the nuclei. Stained samples were mounted
and visualized under a fluorescent microscope (Thermo Fisher Scientific).
