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RNA Interference of RNF31 Enhances ERα Activity in Breast Cancer Cells

This study investigates the impact of RNA interference of RNF31 on ERα activity in breast cancer cells. The results show that decreased RNF31 expression leads to increased ERα activity, suggesting its potential role as a negative regulator of ERα.

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RNA Interference of RNF31 Enhances ERα Activity in Breast Cancer Cells

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  1. Fig. S1 A siControl E2 siER E2 siRNF31 E2 G1: 45.7% S: 26.9% G2-M: 27.4% G1: 70.9% S: 9.9% G2-M: 19.2% G1: 57.1% S: 12.0% G2-M: 30.9% B siControl siRNF31 siRNF31+ ERα plasmid G1: 61.7% S: 13.8% G2-M: 24.9% G1: 57.6% S: 22.6% G2-M: 20.3% G1: 55.4% S: 18.9% G2-M: 26.2%

  2. Fig. S2 A MDAMB175 T47D siControl siControl siRNF31 siRNF31 120 kDa 66 kDa 37 kDa 120 kDa 66 kDa 37 kDa RNF31 ER GAPDH RNF31 ER GAPDH B RNF31 (mRNA) 1.2 1.0 0.8 0.6 0.4 0.2 0 siRNF31 SMARTpool Relative mRNA level siRNF31 oligo 1 siRNF31 oligo 2 siControl RNF31 ER GAPDH 120 kDa 66 kDa 37 kDa

  3. Fig. S3 A B NA NA NA NA Relative mRNA level C D NA NA *** *** *** Relative luciferase activity *** *** *** Relative mRNA level Relative luciferase activity NA

  4. Fig. S4 A IP IgG IP RNF31 B IP: RNF31 IB: ER Input IgG IP RNF31 Input IgG IP RNF31 Input IgG IP RNF31 Input IP IgG IP ER IP: ER IB: RNF31 ER Input ER ER IgG Ig G Ig G C Input IgG IP ERα Input IgG IP ERα Input IgG IP ERα RNF31 RNF31 RNF31 Ig G Ig G Ig G

  5. Fig. S5 nucleus cytosol nucleus cytosol nucleus cytosol nucleus cytosol

  6. Fig. S6 RNF31 ER Merge Merge with nuclear stain

  7. Fig. S7  Fold enrichment over IgG control

  8. Table S1 Real-time PCR primers used in this study

  9. Table S2 RNF31 expression correlates with expression of known ERα target genes Yellow: p-value < 0.05; Red: positive correlation; Green: negative correlation

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