Anti stat3 h 190

The following antibodies were used in this study: anti-β-actin (I–19, 1 : 500), anti-Histone H3 (FL-136, 1 : 500) and anti-STAT3 (H-190, 1 : 1000) antibodies (Santa Cruz Biotechnology); a monoclonal anti-Flag M2 antibody (Sigma-Aldrich, 1 : 1000); anti-EGFR antibody (D38B1, 1 : 1000), anti-phospho-EGFR (Y1173)(53A5, 1 : 1000), anti-Tri-Methyl-Histone H3 (Lys4)(#9727, 1 : 1000), anti-Tri-Methyl-Histone H3 (Lys27)(C36B11, #9733, 1 : 1000), anti-STAT3 (124H6, #9139, 1 : 1000), anti-Acetyl-Histone H3 (Lys27) (#4353, 1 : 1000), anti-NF-κB p65 (#3034, 1 : 1000), and anti-phospho-STAT3 (Tyr705) (D3A7, 1 : 1000) antibodies (Cell Signaling Technology); an anti-ID1 antibody (ab168256, Abcam, 1 : 500); an anti-acetyl-Histone H3 (Lys23) antibody (#07-355, 1 : 1000, Millipore-Upstate); an anti-TRIM24 antibody (#14208-1-AP, 1:500, Proteintech Group). The secondary antibodies were from Vector Laboratories or Jackson ImmunoResearch Laboratories.Peroxidase blocking reagent was from DAKO. AquaBlock was from East Coast Biologics, Inc. Erlotinib was from LC Laboratories. Cell culture media and other reagents were from Invitrogen, Sigma-Aldrich, VWR, or ThermoFisher Scientific.

Different types of treated HLCZ01 cells were lysed in a RIPA buffer supplemented with proteinase inhibitors. 30 μg of proteins were loaded and separated on SDS-PAGE gel; then, the proteins were transferred onto a PVDF membrane, blocked in 5% (w/v) non-fat milk, and incubated with the primary antibodies. Sources of the primary antibodies were anti-STAT3 (H-190) (Santa Cruz, H-190, sc-7179), p-STAT3 (Tyr705) (Santa Cruz, sc-7993), anti-SALL1 (cat# ab130705), anti-SALL2 (Proteintech, cat#12679-1-AP), and anti-SALL4 mAb (Abcam, cat#ab61703). Protein bands were visualized by enhanced chemiluminescence (Milipore).