Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Oct 22;9(11):1081. doi: 10.1038/s41419-018-1125-5

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 3 — a Annexin-V/PI high content microscopy analysis of PC3, DU145 and VCaP cells treated for 24 h with 10 nM scrambled control (SC) siRNA, FLIP(S) specific (FS) siRNA, FLIP(L)-specific (FL) siRNA or siRNA targeting both FLIP(L) and FLIP(S) spliceforms (FT) siRNA followed by 24 h treatment with 1 μM TL32711(TL) + /−10ng/mL TNFα. b Western blot of PARP, cIAP1 and FLIP expression in PC3, DU145 and VCaP cells treated for 24 h with 10 nM scrambled control (SC) siRNA, FLIP(S) specific (FS) siRNA, FLIP(L)-specific (FL) siRNA or siRNA targeting both FLIP(L) and FLIP(S) spliceforms (FT) siRNA followed by 6 h treatment with 1 μM TL32711 in combination with 10 ng/mL TNFα. c, Left – Western blot of FLIP, FADD, RIPK1 and caspase-8 following caspase-8 immunoprecipitation in PC3 cells treated for 24 h with 10 nM Scramble Control (SC), FLIP-Total (FT) or untransfected (−) followed by 3 h with 1 µM TL32711 and 10 ng/mL TNFα; cells were pretreated with 20 µM z-VAD-fmk. Total cell lysate is presented in the panel on the right. d Western blot of FLIP, HDAC1 and HSP90 in cytoplasmic (cC) and nuclear (nN) fractions following 24 h treatment with 10 nM scrambled control (SC) siRNA or siRNA targeting both FLIP(lL) and FLIP(sS) spliceforms (FT). e Western blot of PARP and FLIP expression (Left), and caspase-3/7 activity assay (right) in DU145 cells treated for 1 h with 1 ng/mL Leptomycin-B (Lep), 6 h TL32711(TL) + TNFα or pretreated for 1 h with Leptomycin-B(Lep) followed by 6 h TL32711(TL) + TNFα (Combo). f Cell viability assay of PC3 cells retrovirally infected to stably overexpress empty vector(EV), wild-type FLIP(sS)(FS WT), wild-type FLIP(L) (FL WT) and FADD-binding-deficient F114A mutant FLIP(sS) (FS F114) treated for 72 h with 1 μM TL32711 in combination with 10 ng/mL TNFα (Left). Right: Western blot analysis of parental PC3 and PC3 cells retrovirally infected to stably overexpress empty vector(EV), wild-type FLIP(sS)(FS WT), wild-type FLIP(L)(FL WT) and FADD-binding-deficient F114A mutant FLIP(sS)(FS F114A). *p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001