β tubulin sc 9104

Plumbagin, α-MSH (M4135), arbutin (A4256), kojic acid (K3125), l-DOPA (333786), and tyrosinase (T3824) purified from mushrooms were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against MITF (#12590), p-AKT^S473 (#4060), p-AKT^T308 (#13038), p-CREB (#9198), p-ERK1/2 (#4370), ERK1/2 (#9102), γH2AX (#9718), PARP (#5625), and caspase-3 (#9665) were purchased from Cell Signaling Technology (Danvers, MA, USA). Tyrosinase (sc-7833), and β-tubulin (sc-9104) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

The anti-Tax1 antibodies Lt-4 and Taxy-7 and the anti-Tax2 (GP3738) antibody have been previously described [40 (link)–42 (link)]. Anti-RelA (sc-372), anti-p52 (sc-3786), anti-p38α (sc-535), anti-JNK (sc-571), anti-nucleolin (sc-13057) and β-tubulin (sc-9104) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-phospho-p38 (#9211), anti-phospho-SAPK/JNK (#9251), anti-phospho-Erk1/2 (#9106) and anti-Erk1/2 (#9102) antibodies were purchased from Cell Signaling (Danvers, MA, USA).

Antibodies against MITF (#12590), p-AKT^S473 (#4060), p-CREB (#9398), p-ERK1/2 (#4370), ERK1/2 (#9102), p-MEK (#9154), MEK (#9122), and ERK1/2 inhibitor U0126 were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-Tyrosinase (sc-7833) and β-tubulin (sc-9104) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-TYRP2 (DCT, ab74073) was purchased from Abcam (Cambridge, UK). Zerumbone (Z3902), arbutin (A4256), kojic acid (K3125), α-MSH (M4135), and L-DOPA (333786) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Stock solution of α-Melanocyte-stimulating hormone was prepared in phosphate buffered saline (PBS) prior to treatment. Recombinant human SCF was obtained from R&D systems (Minneapolis, MN, USA) and its stock solution (10 μM) was prepared in PBS. Stock solutions of zerumbone (20 mM), arbutin (1 M), and kojic acid (0.2 M) were prepared in dimethyl sulfoxide (DMSO). Lyophilized Zingiber officinale extract (035-061), isolated by 99% methanol, was obtained from Korea Plant Extract Bank (KPEB) (Daejeon, Korea) and Korea Research Institute of Bioscience and Biotechnology (KRIBB) (Daejeon, Korea). Stock solution of Zingiber officinale extract was prepared in DMSO prior to treatment.

Protein lysate harvested from MCF-7 cells was quantitated with the BCA protein determination kit (Pierce Thermo Scientific, USA). An equal amount (20-30 μg) of protein lysate was subjected to gel electrophoresis on 10% SDS page gels, transferred to nitrocellulose membrane and probed with antibodies to PARP-1 (C2- 10, Trevigen, diluted 1:1000), Pro-Caspase-7 (PRS3467, Sigma, diluted 1:1000), Pro-Caspase-9 (MA1-12562, Thermo Scientific, diluted 1:1000), pH2AX (ADI-KAM- CC255, Enzo Life Sciences, diluted 1:4000), and CETP (PA1-050, ABR, diluted 1:500). β-Tubulin (sc-9104, Santa Cruz Biotechnology, diluted 1:1000) was used as a loading control. Fold change in expression of the serum protein was determined using densitometry analysis of Western blots using ImageJ.

Recombinant TGFβ1 protein (240-B) was purchased from R&D system (Minneapolis, MN, USA). Anti-Flag (F3165) mouse monoclonal antibody and V5-affinity agarose bead (A7345) were purchased from Sigma Aldrich (St. Louis, MO, USA). Anti-ID1 (sc-133104), β-actin (sc-47778) and β-tubulin (sc-9104) mouse monoclonal antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-E-cadherin (610181) and N-cadherin (610921) mouse monoclonal antibodies were obtained from BD Biosciences. Anti-vimentin (CST-5741) rabbit monoclonal antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-PGC1α (ST1202) and TCF4 (H00006925-M04) mouse monoclonal antibody was purchased from Merck Millipore (Billerica, MA, USA) and Abnova (Taipei, Taiwan). Anti-TWIST1 (ab50887) mouse monoclonal and anti-FOXA1 (ab170933) rabbit monoclonal antibodies were purchased from Abcam (Cambridge, MA, USA). 2′,7′-dichlorofuorescin diacetate (DCF-DA, D68883) was purchased from Sigma Aldrich (St. Louis, MO, USA).