Anti gata3 antibody

ChIP assays were performed with the EZ-Magna Chip™ A kit (Millipore, Boston, MA, USA) following the manufacturer's instructions. A total of 1×10⁷ HUVECs co-cultured with A549 conditioned media or control conditioned media were cross-linked with 1% formaldehyde at room temperature for 10 minutes. Sonication was performed on ice to get 200 to1000 bp DNA fragments. The chromatin was then immunoprecipitated with anti-IgG antibody (Millipore, Boston, MA, USA) and anti-GATA3 antibody (Abcam, Cambridge, UK). After reverse cross-linking and DNA purification, DNA from input (1:10 diluted) or immunoprecipitated samples were assayed by semi-quantitative PCR. The primers amplifying the GATA binding motif on the vWF promoter are as follows: sense 5′-TGGGCGGCACCATTGT-3′, and antisense 5′- CATACCTTCCCCTGCAAATGA-3′. The PCR products were analyzed by agarose gel electrophoresis.

IHC was performed as previously described [30 (link)]. Immunohistochemical staining was performed using the standardized labeled streptavidin biotin (LSAB) kit (Dako Cytomation, Carpinteria, CA) according to the manufacturer's instructions. The slides were incubated with rabbit polyclonal anti-GATA3 antibody (1:100 dilution, Abcam, Cambridge, MA, UK), and anti-FGFR1 anti-body (1:100 dilution, Abcam, Cambridge, MA, UK). The slides were evaluated blindly by two independent observers (B.H. And W.T.), and the scoring systems to validate FGFR1, GATA3 expression were described before [30 (link)].