Grem1 antibody

MDA-MB-231 and MTV/TM-011 cells were originally obtained from American Type Culture Collection and Korean Cell Line Bank, respectively. MDA-MB-231 cells were cultured in DMEM (Corning Inc., Corning, NY, USA) containing 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin/streptomycin (Corning Inc.). MTV/TM-011 cells were cultured in RPMI (Corning Inc.) containing 10% FBS and 1% penicillin/streptomycin. Cells were maintained at 37 °C in a humidified atmosphere with 5% CO₂/95% air. Rabbit polyclonal GREM1 antibody (Cat#. 140010) was purchased from Abcam (Cambridge, UK). Recombinant human GREM1 (Cat#. 5190-GR) was obtained from R&D systems (Minneapolis, MN, USA). Anti-MMP13 antibody (Cat#. 515284) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-phospho STAT3 (Cat#. 9145s) and anti-STAT3 (Cat#. 9139s) were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-β-actin antibody (Cat#. A1978) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Stattic (CAS. No. 2798) was purchased from Tocris (Bristol, UK).

MCF-10A, MCF-10A-ras, MDA-MB-453, MDA-MB-468, SKBR3, MCF-7, T47D, and CCD-1068sk cells were originally obtained from American Type Culture Collection, and the BT474 cell line was obtained from Korean Cell Line Bank. The cells were cultured according to the standard procedure and maintained at 37 °C in a humidified atmosphere composed of 5% CO₂/95% air. GREM1 antibody was purchased from Abcam and recombinant human GREM1 was obtained from R&D systems. Recombinant human EGF, anti-Flag antibody and cell linker kits (PKH26 and PKH67) were purchased from Sigma-Aldrich. Anti-ERRα, anti-p-EGFR/EGFR, anti-p-Akt/Akt, anti-p-ERK/ERK antibodies and erlotinib were obtained from Cell Signalling Technology. Expression plasmids of GREM1, ERRα, Flag-only, Flag-EGFR, and Flag-BMP2 were purchased from Sino Biological Inc. Fc-IgG1 and Fc-GREM1 were provided by ACROBiosystems. The lentiviral GREM1 clone was obtained from Genecopoeia. 3xERRE-luciferase, pcDNA4-myc-PGC-1α, EGFR-WT, EGFR-ECD and EGFR-ICD were provided by Addgene. XCT790, LY294002 and U0126 were purchased from Tocris.