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

. 2022 Oct 6;30(1):208–220. doi: 10.1038/s41418-022-01066-0

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

© The Author(s) 2022

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. 2 — a Representative flow cytometry histograms of a caspase-10 activity assay performed on living cells at indicated days (N = 4). b (Left panels) Western blots detection of caspase-10 and caspase-8 performed on sorted live cells (PI/annexin-V negative) at indicated time of the CD36 culture for expression of KIT/GPA: day 3: KIT⁺GPA^low, day 6: KIT^+/lowGPA^int, day 8: KIT^lowGPA^high. (*) Caspase-10 activated forms p29 kDa and p17 kDa. (Right panels) Western blots performed on day 5 of the CD36 culture performed at several EPO concentrations in the presence or not of the Fas-L mimetic monoclonal antibody CH11. *Fully activated caspase-8 (P10 kDa), fully activated caspase-10 (P17 kDa) in addition to p29kDa. (N = 3). c Representative flow cytometry histograms of a caspase-10 activity assay at indicated days, performed on living erythroid cells cultured from bone marrow CD34 + cells (N = 3). d A schematic representation of the different BID fragments generated by caspase-8 and -10 cleavage. Cleavage at position D60 exposes a glycine to N-myristoylation (M), generating Myr-P15-tBID. A putative caspase-10 cleavage site at position D38 is indicated. e Representative western blot analysis of BID performed on live, sorted cells (PI/annexin-V-negative) at different time points of the culture for expression of KIT/GPA: day 2: KIT⁺GPA^low, day 6: KIT^+/-GPA^int, day 8: KIT^lowGPA^high (N = 6). Cleaved forms P18 and P13 are indicated. f Representative western blot analyses of BID performed day 7 on: (+) live sorted cells (PI/annexin-V-negative) or (TL) total lysates of the same culture, expressing wild-type P22-BID (WT), P22-BID-D60A and P22BID-D75A. (P18-tBID (•), P15-tBID (✦), P13-tBID (✶)). (N = 4). g Western blot anti-BID and anti-caspase-10 performed on day 7 of sh1CASP10-GFP + and control transduced cells culture. h Semi-tryptic analysis of the mass spectrometry data (using Mascot) extracted from the 18 kDa gel area (Mascot score for the upper spectrum: 54) and the 13 kDa gel area (Mascot score for the lower spectrum: 30).