Identification of PLXDC1 and PLXDC2 as the transmembrane receptors for the multifunctional factor PEDF
- Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, United States
- Jules Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, United States
- University of Oklahoma Health Sciences Center, United States
Figures
Figure 1 with 1 supplement
The binding of PEDF to PLXDC1 and PLXDC2 on cell surface.
Upper panel: Binding of biotinylated PEDF to control HEK293 cells (A) or HEK293 cells transfected with PLXDC1 (B), PLXDC2 (C), intracellular domain deleted PLXDC2 (D, PLXDC2-dC), extracellular …
Figure 1—figure supplement 1
PLXDC1 and PLXDC2 schematic diagrams and alignment showing the definitions of domains.
(A) Schematic diagrams of domains in PLXDC1 (red) and PLXDC2 (orange). (B) Alignment of human (h) and mouse (m) PLXDC1 and PLXDC2. Junctions of domains are indicated. Identical regions are …
Figure 2 with 1 supplement
The roles of PLXDC1 and PLXDC2 in PEDF-induced IL-10 secretion by macrophage cell RAW267.4.
(A) siRNA-mediated knockdown of PLXDC1 or PLXDC2 substantially suppresses PEDF-stimulated IL-10 secretion. Activity of control transfected cells with PEDF treatment is defined as 1. ** = p < 0.01. (B…
Figure 2—figure supplement 1
Unbiased screening for effective siRNAs that knock down PLXDC1 or PLXDC2 expression in macrophage cell RAW267.4.
The definition and source of each siRNA is described in ‘Materials and methods’.
Figure 3 with 1 supplement
The PLXDC2 dependence of PEDF's effect on endothelial cell SVEC4-10.
(A) PEDF promoted-cell death of SVEC4-10 cells is suppressed by siRNA-mediated knockdown of PLXDC2, but not PLXDC1. Survival of control siRNA tranfected cells without PEDF treatment is defined as 1. …
Figure 3—figure supplement 1
Unbiased screening for effective siRNAs that knock down PLXDC1 or PLXDC2 expression in endothelial cell SVEC4-10.
The definition and source of each siRNA is described in ‘Materials and methods’.
Figure 4 with 1 supplement
PEDF's neurotrophic effect on 661W cells depends on PLXDC1.
(A) PEDF treatment protects 661W cells against hydrogen peroxide-mediated oxidative damage. siRNA-mediated knockdown of PLXDC1, but not PLXDC2 abolishes the protective effect of PEDF. The survival …
Figure 4—figure supplement 1
Unbiased screening for effective siRNAs that knock down PLXDC1 or PLXDC2 expression in neuronal cell 661W.
The definition and source of each siRNA is described in ‘Materials and methods’.
Figure 5 with 1 supplement
Receptor oligomerization.
(A) PLXDC1 deletion series with a Rim tag following the N-terminal secretion signal were purified together with HA-tagged full length PLXDC1 by purifying the Rim tag (diagrams on the left). Anti-Rim …
Figure 5—figure supplement 1
PLXDC1 and PLXDC2 preferentially form homooligomers.
(A) Schematic diagram of experimental design to compare homooligomer and heterooligomer formation of PLXDC1 is shown on the left. Purification of the Rim tag is indicated by an antibody symbol. …
Figure 6
Structure/function and real-time analysis of PEDF/receptor interaction.
(A) PEDF with HA tag after the N-terminal secretion signal is copurified with different deletion mutants of PLXDC1, which are all tagged with a Rim tag following the N-terminal secretion signal …
Figure 7
Crosslinking of the terminal cysteine of PLXDC1 prevents PEDF's effect on receptor dimerization.
(A) Schematic diagrams of the experimental design. ‘C’ indicates the cysteine residue located at the C-terminus of PLXDC1. ‘S’ indicates mutation of this residue to serine. (B and C) Crosslinking of …
