Hydroquinone regulates hemeoxygenase-1 expression via modulation of Src kinase activity through thiolation of cysteine residues

In vitro and in vivo anti-inflammatory activity of Phyllanthus acidus methanolic extract

“…RAW264.7 cells, a BALB/c-derived murine macrophage cell line (TIB-71); U937, a human promonocytic cell line (CRL-1593.2); and HEK293 cells, a human embryonic kidney cell line (CRL-1573) were purchased from ATCC (Rockville, MD, USA). Luciferase constructs containing binding sites for NF-κB and epitope-tagged signalling expression constructs (FLAG-MyD88, HA-Src, and Myc-Syk) were used as previously reported (Byeon et al, 2013). All other chemicals were purchased from Sigma.…”

“…After treatment with Pa-ME, cells were incubated for 12 h and luciferase assays were performed using the Luciferase Assay System (Promega, Madison, WI, USA) as previously reported (Back et al, 2013). HEK293 cells (1 Â 10 7 cells/ml) were transfected with HA-Src or Myc-Syk for 48 h as previously reported (Byeon et al, 2013). Before harvesting, the cells were treated with Pa-ME for 12 h. Lysates of HA-Src-overexpressing or Myc-Syk-overexpressing cells were analysed by immunoblotting to determine the levels of total and phospho-form Src, Syk, p85, AKT, and β-actin.…”

TGFβ1 rapidly activates Src through a non-canonical redox signaling mechanism

“…To identify the cysteine residue(s) in human Src involved in Src activation by TGF-β, cysteine residues at 248, 277, 490, or 501 were replaced individually with alanine, using site-directed mutation. These cysteine residues were selected because of their conservation among Src kinase family members across species (39), or their reported sensitivity to redox modification (35,40,50). As shown in Figure 6, the phosphorylation of Src at Tyr419 and Tyr530 were affected in different ways by the cysteine/alanine mutation.…”

“…On the other hand, there are also reports that some cysteine residues in Src might be involved in the suppression of Src activity. For instance, Cys277 was found to be responsible for oxidant-mediated Src homodimerization and inactivation in an in vitro assay (40), and mutation of Cys403 and Cys486 increased Src activity (50). Our data support that cysteine residue 248, 277, 490 and 501 of human Src are essential for Src activation by TGF-β and are potential targets of oxidative modification.…”

PDK1 in NF-κB signaling is a target of Xanthium strumarium methanolic extract-mediated anti-inflammatory activities

“…CRL-1573), were purchased from ATCC (Rockville, MD, USA). Luciferase constructs containing binding sites for NF-κB and PDK1 were used as reported previously (Hossen et al, 2015b) (Byeon et al, 2013). All other chemicals were purchased from Sigma.…”