Journal: Oncology Letters
Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells
doi: 10.3892/ol.2025.15155
Figure Lengend Snippet: RT-qPCR validation of changes in expression of a subset of genes following BLID knockdown. RT-qPCR analysis of several genes in (A) MCF-7 and (B) MDA-MB-231 cells. β-Actin served as the internal control. (A) The y-axis title of the middle graph is identical to the y-axis title of the left graph. (B) The y-axis title of the middle-left graph is identical to the y-axis title of the far-left graph. *P<0.05 and **P<0.01 (BLID shRNA vs. the corresponding Scr shRNA group), n=3. shRNA, short hairpin RNA; Scr, scramble control; Ctl, control; RT-qPCR, reverse transcription-quantitative PCR; BLID, BH-3 like motif containing inducer of cell death.
Article Snippet: The BLID shRNA constructs used were as follows: Construct 1, shRNA TRCN0000265539 (RefSeq- NM_001001786.1 –656s21c1) forward sequence, 5′-CCGGGGACAGATTTCGCCCATTATTCTCGAGAATAATGGGCGAAATCTGTCCTTTTTG-3′ and reverse sequence, 5′-AATTCAAAAAGGACAGATTTCGCCCATTATTCTCGAGAATAATGGGCGAAATCTGTCC-3′; construct 2, shRNA TRCN0000254449 ( NM_001001786.1 –516s21c1) forward sequence, 5′-CCGGTCTGCCATGAAGCGGAATGTTCTCGAGAACATTCCGCTTCATGGCAGATTTTTG-3′ and reverse sequence, 5′-AATTCAAAAATCTGCCATGAAGCGGAATGTTCTCGAGAACATTCCGCTTCATGGCAGA-3′; construct 3, shRNA TRCN0000254448 (RefSeq- NM_001001786.1 –566s21c1) forward sequence, 5′-CCGGTTTAACCAGGATACAAGTTACCTCGAGGTAACTTGTATCCTGGTTAAATTTTTG-3′ and reverse sequence, 5′-AATTCAAAAATTTAACCAGGATACAAGTTACCTCGAGGTAACTTGTATCCTGGTTAAA-3′; construct 4, shRNA TRCN0000254447 (RefSeq- NM_001001786.1 –387s21c1) forward sequence,5′-CCGGGCCTCTGGCAGTTCCATTTATCTCGAGATAAATGGAACTGCCAGAGGCTTTTTG-3′ and reverse sequence, 5′-AATTCAAAAAGCCTCTGGCAGTTCCATTTATCTCGAGATAAATGGAACTGCCAGAGGC-3′; and construct 5, shRNA TRCN0000254446 (RefSeq- NM_001001786.1 –449s21c1) forward sequence, 5′-CCGGTTCCAACAAAGAACCTATGTTCTCGAGAACATAGGTTCTTTGTTGGAATTTTTG-3′ and reverse sequence, 5′-AATTCAAAAATTCCAACAAAGAACCTATGTTCTCGAGAACATAGGTTCTTTGTTGGAA-3′. pLKO.1-puro scramble shRNA (Addgene: scramble shRNA Sequences; Addgene plasmid# 1864 RRID:Addgene_1864; http://n2t.net/addgene:1864 ) was used as the negative control.
Techniques: Quantitative RT-PCR, Biomarker Discovery, Expressing, Knockdown, Control, shRNA, Reverse Transcription, Real-time Polymerase Chain Reaction
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