Anti hif 1α or rabbit igg control

Chromatin immunoprecipitation (ChIP) assays were performed as described previously.^{49 (link), 50 (link)} The cells were crosslinked with final concentration 1% formaldehyde for 10 min at room temperature. Then 125 mM glycine was added to quench unreacted formaldehyde. The cells were gathered and sonicated to make DNA fragments with a size range of 200 to 1000 bp. The cell extracts were immunoprecipitated using 2 μg anti-HIF-1α or rabbit IgG control (Abcam, Cambridge, UK) for each sample suspended in 450 μl ChIP dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8.1, 167 mM NaCl) purchased from Cell Signaling Technology (Cat #20–153, Danvers, MA, USA). For all ChIP experiments, PCR analysis were performed by using multiple sets of primers spanning the transcription factor binding site on hBAFF gene promoter.

ChIP assays were performed, as described previously [67 (link),68 (link),69 (link)]. Cells were crosslinked with a final concentration 1% formaldehyde for 10 min at room temperature. Then, 125 mM of glycine was added to quench unreacted formaldehyde. Cells were collected and sonicated to make DNA fragments with a size range of 200 to 1000 bp. Cell extracts were immune-precipitated using 1 μg anti-HIF-1α or rabbit IgG control (Abcam, Cambridge, UK) for each sample suspended in 450 μL ChIP dilution buffer (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8.1, 167 mM NaCl) purchased from Cell signaling Technology (Cat# 20-153, Danvers, MA, USA). For all ChIP experiments, PCR analyses were performed by using multiple sets of primers spanning the transcription factor binding site on the NPHP3 gene promoter.