The blotted membranes were blocked with 5% non-fat dry milk (w/v) in Tris-buffered saline with 0.1% Tween-20 (TBS-T) for 1 h at room temperature and then incubated with the following primary antibodies overnight at 4C: anti-Smad2 (1:1,000 dilution, Cell Signaling Technology), anti-phospho-Smad2 (1:1,000 dilution, Cell Signaling Technology), anti-Smad3 (1:1,000 dilution, Cell Signaling Technology), anti-phospho-Smad3 (1:1,000 dilution, Cell Signaling Technology), anti-TGF-1 (1:1000 dilution, Abcam), anti-IGF-1 (1:500 dilution, Abcam), TIMP-1 (1:500 dilution, Santa Cruz), MMP-9 (1:1000 dilution, R&D), fibronectin (1:500 dilution, Santa Cruz), E-cadherin (1:2000 dilution, CST), vimentin (1:2000 dilution, CST), and -SMA (1:2000 dilution, CST)
It may help reduce oxidative stress that fuels dull tone and pigmentation
Notably, glucobrassicina major indole glucosinolateand proline, an osmoprotectant, were strongly enriched in the stress + FFA group, reflecting enhanced secondary metabolism and osmotic adjustment
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