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Journal: Nucleic Acids Research
Article Title: A cautionary tale of sense-antisense gene pairs: independent regulation despite inverse correlation of expression
doi: 10.1093/nar/gkx952
Figure Lengend Snippet: NOP14-AS1 : an antisense transcript induced upon DNA damage. ( A ) Microarray analysis heat map identifying lncRNAs and mRNAs differentially expressed in A549 (left panel) and HepG2 (right panel) cells treated with 50 μM Etoposide (ETO)/50 μM Cisplatin (CIS)/20 μM Bleomycin (BLEO) or vehicle control DMSO for 8 h. ( B ): A549/HepG2/NCI-H460/HEK293/MCF7 cells were treated with 50 μM Etoposide (ETO)/50 μM Cisplatin (CIS)/20 μM Bleomycin (BLEO)/1 μM DOXO/50 μM Carboplatin (CARBO) or vehicle control DMSO for 8 h. RT-qPCR results for NOP14-AS1 normalized to Cyclophilin A and DMSO control. Error bars represent SEM ( n ≥ 3). * P < 0.05; ** P < 0.01; *** P < 0.001 compared to DMSO control, unpaired two-sided t -test. ( C ) A549/NCI-H460/HepG2 cells were treated with either 10 μM Nutlin-3 or vehicle control DMSO for 24 h. RT-qPCR results for NOP14-AS1 normalized to Cyclophilin A and DMSO controls. Error bars represent SEM ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001 compared to DMSO controls, unpaired two-sided t -test. ( D ) HCT116 TP53 WT/TP53 Null cells were treated with 50 μM Etoposide (ETO)/1 μM DOXO/10 μM Nutlin-3 or vehicle control DMSO for 12 h. RT-qPCR results for NOP14-AS1 normalized to Cyclophilin A and DMSO controls. Error bars represent SEM ( n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001 compared to DMSO controls, unpaired two-sided t -test. ( E ) NCI-H460 cells were transfected with either one of the two indicated siPOOLs against TP53 or siPOOL Control. These were then treated with either 1 μM DOXO or vehicle control DMSO for 24 h. Upper panel: RT-qPCR results for NOP14-AS1 normalized to Cyclophilin A and siPOOL Control + DMSO control. Error bars represent SEM ( n = 5). * P < 0.05; ** P < 0.01; *** P < 0.001 compared to siPOOL Control, unpaired two-sided t -test. Lower panel: western blot results for TP53. GAPDH was used as a loading control.
Article Snippet: Etoposide (Topoisomerase II inhibitor, induces double-strand breaks in genomic DNA) (33419–42-0, Cayman Chemical), Cisplatin (forms intrastrand cross-links with purine bases in genomic DNA) (CAS 15663–27-1, Merck Millipore),
Techniques: Microarray, Quantitative RT-PCR, Transfection, Western Blot