3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt

GHK and GHK-Cu were purchased from McBiotec (Nanjing, China). The ratio of GHK to Cu is 2:1 according to manufacturer data. Copper (II) chloride, 99%, and copper (II) acetate monohydrate, ACS reagent, ≥98%, powder were purchased from Sigma-Aldrich (St. Louis, USA). Phosphate buffered saline (PBS) (pH 7.4, 10×) was obtained from Vivantis (Malaysia). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum, random primers and SYBR safe DNA gel stain were supplied by Invitrogen, Life Technologies (USA). Trypsin and penicillin/streptomycin solution were obtained from PAN-Biotech GmbH (Germany). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were acquired from MP Biomedicals (Illkirch, France). RNeasy Mini Kit and QuantiFast SYBR Green PCR kit were purchased from Qiagen (Germany). random primers and avian myeloblastosis virus reverse transcriptase were purchased from Promega (Madison, Wisconsin, USA). Human interleukin 1 alpha and interleukin 8 enzyme-linked immunosorbent assay (ELISA) kits were purchased from Biolegend (San Diego, CA, USA). Human Fos-related antigen 1 (FOSL1) and Heat Shock 70 kDa Protein 1A (HSPA1A) ELISA kits were purchased prom MyBioSource (San Diego, CA, USA). All chemicals were used as supplied.

Antibodies against E-cadherin, N-cadherin, and Matrigel basement matrix were purchased from BD Biosciences (San Jose, CA, USA). Antibodies against IGFBP-3, vimentin, myc, glutathione S-transferase (GST), His-probe, HA-probe, OctA-probe, IGF-1R, ubiquitin, and actin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against E-cadherin, N-cadherin, and tubulin were purchased from Cell Signaling Technology (Danvers, MA, USA). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from GeneTex (Irvine, CA, USA). Fluorochrome (Alexa Fluor 488, Alexa Fluor 546, or Alexa Fluor 594)-conjugated secondary antibodies were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Human recombinant IGFBP-3 was purchased from R&D Systems (Minneapolis, MN, USA). Human recombinant IGFBP-3 protein used or evaluation of cellular uptake of extracellular IGFBP-3 was kindly provided by Insmed Inc. (Glen Allen, VA, USA) [22 (link)]. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from MP Biomedicals (Santa Ana, CA, USA). G418 was purchased from Enzo Life Sciences (Farmingdale, NY, USA). Crystal violet, a mouse monoclonal anti-vimentin antibody, and additional chemicals unless otherwise indicated were purchased from Sigma-Aldrich (St. Louis, MO, USA).

2-Bromo-2-methylpropionyl bromide, triethylamine (TEA), N,N′-dicyclohexylcarbodiimide (DCC), 4-dimethylaminopyridine (DMAP), and copper(I) chloride (CuCl) were purchased from Alfa Aesar (Heysham, UK). Polyethylene glycol monomethyl ether (mPEG, Mw = 2000 g/mol) was obtained from TCI (Tokyo, Japan). N,N,N′,N″,N″-Pentamethyldiethylenetriamine (PMDETA), 2-hydroxyethyl disulfide, o-nitrobenzyl alcohol and succinic anhydride were purchased from Sigma-Aldrich (St. Louis, MO, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) was from MP Biomedicals (Santa Ana, CA, USA). Phosphate buffer saline (PBS), Dulbecco’s Modified Eagle Medium (DMEM), Roswell Park Memorial Institute (RPMI) 1640 Medium, trypsin–EDTA, and fetal bovine serum (FBS) were purchased from Invitrogen (Waltham, MA, USA). Doxorubicin hydrochloride (DOX·HCl) was purchased from Combi-Blocks (San Diego, CA, USA).