Journal: Nucleic Acids Research

Article Title: Coactivators enable glucocorticoid receptor recruitment to fine-tune estrogen receptor transcriptional responses

doi: 10.1093/nar/gkt100

Figure Lengend Snippet: MED14, SRC-2 and SRC-3 mediate the recruitment of GR to the PRL array. ( A ) Heat map showing effects of siRNA knockdown of multiple factors on features of the PRL array system, including GR loading, active RNA polymerase II loading and ER loading. ( B ) Quantification of GR nuclear intensity (normalized to control siRNA) after 72 h siRNA knockdown of GR, MED14 and SRC-2/-3 in HeLa-PRL:GFP-ERα cells treated with 10 nM E2 + 100 nM Dex for 30 min. ( C ) Quantification of MED14 nuclear intensity (normalized to control siRNA). ( D ) Quantification of SRC-2 nuclear intensity (normalized to control siRNA) treated as described earlier in the text. ( E ) Quantification of SRC-3 nuclear intensity (normalized to control siRNA). ( F ) Quantification of GR loading (normalized to control siRNA) after 72 h siRNA knockdown of GR, MED14 or SRC-2/-3 in HeLa-PRL:GFP-ERα cells treated with 10 nM E2 + 100 nM Dex for 30 min. ( G ) Quantification of MED14 loading (normalized to control siRNA) after 72 h siRNA knockdown of MED14, or SRC-2/-3 in HeLa-PRL:GFP-ERα cells treated with 10 nM E2 + 100 nM Dex for 30 min. ( H ) Quantification of SRC-2 loading (normalized to control siRNA) after 72 h siRNA knockdown of MED14, or SRC-2/-3 in HeLa-PRL:GFP-ERα cells treated with 10 nM E2 + 100 nM Dex for 30 min. ( I ) Quantification of SRC-3 loading (normalized to control siRNA) after 72 h siRNA knockdown of MED14, or SRC-2/-3 in HeLa-PRL:GFP-ERα cells treated with 10 nM E2 + 100 nM Dex for 30 min (* P < 0.05; ** P < 0.005; *** P < 0.0005).

Article Snippet: The antibodies used were anti-GR (GeneTex GTX101120), RNA polymerase II (ABCAM ab5408), PPARγ (ABCAM ab52270), ERα (Millipore 04-820), Ser5-phospho RNA polymerase II (ABCAM ab5401), MED14 (Bethyl A301-044 A), SRC-2 (BD Transduction Labs # 610985) and SRC-3 (BD Transduction Labs # 611105).

Techniques: