1 / 1

Investigating the Impact of PTB Knockdown on BAG-1 Isoform Expression in HeLa Cells

This study examines the effect of PTB knockdown on BAG-1 isoform expression in HeLa cells. Cells were transfected with siRNAs targeting PTB and a control sequence, followed by transfection with either the di-cistronic plasmid pRBagF or control plasmid pRF. Luciferase activity was measured using a Dual-Luciferase reporter assay system, and Western analysis was performed to assess PTB and BAG-1 expression levels, with actin as a loading control. Results indicate a significant reduction in BAG-1 p36 isoform levels and IRES activity upon PTB knockdown.

shika
Télécharger la présentation

Investigating the Impact of PTB Knockdown on BAG-1 Isoform Expression in HeLa Cells

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. C2 P1 PTB p50 BAG-1 p46 p36 Actin 1.2 1 0.8 p1 0.6 Relative luciferase activity c2 0.4 0.2 0 pRF pRBagF Supplementary data 1. HeLa cells were transfected with siRNAs directed against PTB P1 (AAC UUC CAU CAU UCC AGA GAA) or a control sequence and C2 (AAG GUC CGG CUC CCC CAA AUG) as described previously (Mitchell et al 2005). Cells were then transfected with the di-cistronic plasmid pRBagF (as described; Coldwell et al 2000) or the control plasmid pRF. Lysates were prepared from transfected cells and luciferase activities were measured and calculated as previously described (Mitchell et al., 2003) using a Dual-Luciferase reporter assay system (Promega). Western analysis was performed on these samples to detect expression of PTB, and BAG-1 whilst actin was used as a loading control. It can be seen that when PTB levels are reduced there is a specific reduction in the level of the p36 isoform of BAG-1 and a corresponding reduction in BAG-1 IRES activity.

More Related