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

. 2021 Apr 5;5(7):1922–1932. doi: 10.1182/bloodadvances.2020002402

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

© 2021 by The American Society of Hematology

PMC Copyright notice

Figure 3. — Zinc binding by CALR^del52is required for homomultimerization and enables MPL activation. (A) Schematic depicting co-immunoprecipitation assay for detecting FLAG-tagged and V5-tagged CALR^del52 multimers. (B) FLAG-pulldown assays demonstrating CALR^del52 lectin variants retain ability to bind to V5-CALR^del52. (C) FLAG-pulldown assays demonstrating 2His- and 3His-CALR^del52 are compromised in ability to bind V5-CALR^del52 compared with 1His-CALR^del52. (D) FLAG-pulldown assays of cells transfected with FLAG- and V5-tagged CALR^WT (lane 1), CALR^del52 (lane 2), CALR^del52 D135A lectin variant (lane 3), or 3His-CALR^del52 variant (lane 4) demonstrating that CALR^del52 and CALR^del52 D135A are capable of homomultimerization, but not CALR^WT and 3His-CALR^del52. (E) Affinity chromatography demonstrating 2His- and 3His-CALR^del52 bind less efficiently to zinc resin compared with 1His-CALR^del52. (F) Schematic depicting co-immunoprecipitation assay for detecting heteromeric complexes comprising FLAG-tagged and V5-tagged CALR^del52 proteins and MPL. (G) FLAG-pulldown assays demonstrating zinc chelator TPEN treatment disrupts CALR^del52 multimerization and MPL binding in 293T cells.