Journal: Cell reports

Article Title: MeCP2 represses the activity of topoisomerase IIβ in long neuronal genes

doi: 10.1016/j.celrep.2023.113538

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-Topo II-beta (H-286) [secondary] , Santa Cruz , Cat# sc-13059; RRID: AB_2205866.

Techniques: Virus, Recombinant, Mass Spectrometry, Bisulfite Sequencing, Mutagenesis, Derivative Assay, Software

Journal: Cancers

Article Title: A Double-Negative Feedback Interaction between miR-21 and PPAR-α in Clear Renal Cell Carcinoma

doi: 10.3390/cancers14030795

Figure Lengend Snippet: PPAR-α expression and/or activation decreases AP-1 and NF-κB transcriptional activity and binding to miR-21 promoter. ( A ) ACHN and 786-O cells were co-transfected with pSG5-EV (empty vector) or pSG5-PPAR-α expression vectors and κB-Luc or AP1-Luc synthetic promoters. At 24 h after transfection, the cells were incubated with 0 or 400 nM of GW7647. At 48 h after transfection, luciferase activity was measured. Luciferase activity in cells transfected with pSG5-EV was set as 1. ( B , C ) Soluble chromatin from the ACHN and 786-O cells transfected with pSG5-EV or pSG5-PPAR-α expression vectors was immunoprecipitated with control immunoglobulin G (IgG) and anti-c-jun ( B ) or anti-NF-κB p65 antibodies ( C ). The precipitated DNA samples were amplified by PCR with pairs of primers flanking the binding site regions for AP-1 and NF-κB. The results were expressed as the percentage of input. The values obtained with the empty vector were referred to as 1. The values are means ± SEM and represent at least three separate experiments (* p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: The specific antibodies used were: anti-c-jun (D) and anti-NF-κB p65 (H-286) (Santa Cruz Biotechnology, Heidelberg, Germany). qPCR was performed using the SsoFast EvaGreen supermix (Bio-Rad) and the following primers: AP-1 Forward: 5′-TAAGGATGACGCACAGATTGTC-3′; AP-1 Reverse: 5′-TCAGAAGTCCCACATTTATCACC-3′; NF-κB Forward: 5′-GGAGTGGATGGGTTCTGCCTTA-3′; and NF-κB Reverse: 5′-CAAGGTGGATTGCATCGAGG-3′.

Techniques: Expressing, Activation Assay, Activity Assay, Binding Assay, Transfection, Plasmid Preparation, Incubation, Luciferase, Immunoprecipitation, Control, Amplification