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a The expression of SERPINA3 is positively correlated with ESR1 in breast cancer (retrieved from cBioPortal). b SERPINA3 exhibits significant higher expression in ER+ breast cancer ( n = 7247) than ER− subtype ( n = 2710) (retrieved from bc-GenExMiner v4.9). c QPCR and d Western blot results revealed a decrease <t>of</t> <t>ERα</t> expression in LTED cells, when compared with their parental cells. e The expression of ESR1 could be effectively inhibited by the siRNAs in MCF-7 and T47D cells. f QPCR and g western blot results showed that siRNA knockdown of ESR1 in MCF-7 and T47D effectively downregulated SERPINA3 expression. GAPDH and SERPINA3 were blotted from different gels for their close molecular weight. h SERPINA3 mRNA expression decreased with extended estrogen deprivation incubation in MCF-7 and T47D. i The details of the predicted ERα binding sequence in SERPINA3 promoter obtained from JASPAR. j Comparison of ERα binding motif and the predicted ERα binding sequence in SERPINA3 promoter. k Dual luciferase reporter assay verified the physical interaction of ERα and SERPINA3 promoter at −462 to −446 bp. The promoter plasmid or its mutant type were co-transfected with <t>pCDH</t> empty vector or ESR1-expressing plasmid as indicated. Luciferase assays were then performed. pCDH-Vector: pCDH-CMV-MCS-EF1-puro; pCDH-ESR1: pCDH-CMV-MCS-EF1-puro-ESR1. Data are representative of n = 3 biologically independent experiments and presented as mean ± SD; statistical significance is determined by unpaired Student’s t test ( p < 0.05 *, p < 0.01 **, p < 0.001 ***).
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a The expression of SERPINA3 is positively correlated with ESR1 in breast cancer (retrieved from cBioPortal). b SERPINA3 exhibits significant higher expression in ER+ breast cancer ( n = 7247) than ER− subtype ( n = 2710) (retrieved from bc-GenExMiner v4.9). c QPCR and d Western blot results revealed a decrease of ERα expression in LTED cells, when compared with their parental cells. e The expression of ESR1 could be effectively inhibited by the siRNAs in MCF-7 and T47D cells. f QPCR and g western blot results showed that siRNA knockdown of ESR1 in MCF-7 and T47D effectively downregulated SERPINA3 expression. GAPDH and SERPINA3 were blotted from different gels for their close molecular weight. h SERPINA3 mRNA expression decreased with extended estrogen deprivation incubation in MCF-7 and T47D. i The details of the predicted ERα binding sequence in SERPINA3 promoter obtained from JASPAR. j Comparison of ERα binding motif and the predicted ERα binding sequence in SERPINA3 promoter. k Dual luciferase reporter assay verified the physical interaction of ERα and SERPINA3 promoter at −462 to −446 bp. The promoter plasmid or its mutant type were co-transfected with pCDH empty vector or ESR1-expressing plasmid as indicated. Luciferase assays were then performed. pCDH-Vector: pCDH-CMV-MCS-EF1-puro; pCDH-ESR1: pCDH-CMV-MCS-EF1-puro-ESR1. Data are representative of n = 3 biologically independent experiments and presented as mean ± SD; statistical significance is determined by unpaired Student’s t test ( p < 0.05 *, p < 0.01 **, p < 0.001 ***).

Journal: Communications Biology

Article Title: SERPINA3-ANKRD11-HDAC3 pathway induced aromatase inhibitor resistance in breast cancer can be reversed by HDAC3 inhibition

doi: 10.1038/s42003-023-05065-w

Figure Lengend Snippet: a The expression of SERPINA3 is positively correlated with ESR1 in breast cancer (retrieved from cBioPortal). b SERPINA3 exhibits significant higher expression in ER+ breast cancer ( n = 7247) than ER− subtype ( n = 2710) (retrieved from bc-GenExMiner v4.9). c QPCR and d Western blot results revealed a decrease of ERα expression in LTED cells, when compared with their parental cells. e The expression of ESR1 could be effectively inhibited by the siRNAs in MCF-7 and T47D cells. f QPCR and g western blot results showed that siRNA knockdown of ESR1 in MCF-7 and T47D effectively downregulated SERPINA3 expression. GAPDH and SERPINA3 were blotted from different gels for their close molecular weight. h SERPINA3 mRNA expression decreased with extended estrogen deprivation incubation in MCF-7 and T47D. i The details of the predicted ERα binding sequence in SERPINA3 promoter obtained from JASPAR. j Comparison of ERα binding motif and the predicted ERα binding sequence in SERPINA3 promoter. k Dual luciferase reporter assay verified the physical interaction of ERα and SERPINA3 promoter at −462 to −446 bp. The promoter plasmid or its mutant type were co-transfected with pCDH empty vector or ESR1-expressing plasmid as indicated. Luciferase assays were then performed. pCDH-Vector: pCDH-CMV-MCS-EF1-puro; pCDH-ESR1: pCDH-CMV-MCS-EF1-puro-ESR1. Data are representative of n = 3 biologically independent experiments and presented as mean ± SD; statistical significance is determined by unpaired Student’s t test ( p < 0.05 *, p < 0.01 **, p < 0.001 ***).

Article Snippet: ERα over-expressing plasmids pCDH-CMV-MCS-EF1-PURO-ESR1 and its empty vector pCDH were purchased from OriGene (Rockville, MD).

Techniques: Expressing, Western Blot, Knockdown, Molecular Weight, Incubation, Binding Assay, Sequencing, Comparison, Luciferase, Reporter Assay, Plasmid Preparation, Mutagenesis, Transfection