|
|
Blood, Vol. 113, Issue 2, 438-446, January 8, 2009
A functional folate receptor is induced during macrophage activation and can be used to target drugs to activated macrophages
Blood Xia et al.
113: 438
Supplemental materials for: Xia et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 58.3 KB)
- Figure S1. Folate receptors are expressed on a sub-population of thioglycollate-elicited and bacteria-activated macrophages but not on resident peritoneal macrophages (JPG, 95 KB)
-
(A) Representative dot plots of cells show that thioglycollate elicited macrophages express folate receptor. The percentage of macrophages co-stained with the F4/80 macrophage marker is illustrated. (B) Co-incubation with 100 fold excess folic acid blocks folate-FITC binding to the thioglycollate-elicited FR+ macrophage population. (C) Bacteria activated macrophages express folate receptor. (D) Competition study on bacteria-activated macrophages with 100 fold excess folic acid shows that folate-FITC binding is folate receptor specific. (E) Resident peritoneal macrophages do not express or express low folate receptor and (F) shows the competition study of the same population of macrophages. Experiments were repeated three times and the representative dotplot data were shown.
- Figure S2. FR+ macrophages exhibit a more activated morphology than FR− macrophages (JPG, 37.4 KB)
-
Peritoneal macrophages isolated from mice injected with (A) sterile PBS (B) thioglycollate or (C) Pseudomonas aeruginosa, were imaged by confocal fluorescence microscopy (left panels) and brightfield microscopy (right panels) after labeling with folate-rhodamine. (Scale bar: 20 μm).
- Figure S3. The co-expression of TNF-α and FR (JPG, 28 KB)
-
Three color flow cytometry experiments were performed. Peritoneal cells were simultaneously labeled with folate-Oregon green, anti-F4/80 and anti–TNF-α. Experiments were repeated three times and the representative dotplot data were shown.
- Figure S4. Characterization of FR-expressing peritoneal macrophages (JPG, 107 KB)
-
Three color flow cytometry experiments were performed. Peritoneal cells were simultaneously labeled with folate-FITC, anti-F4/80 and one of antibodies against mouse mannose receptor and CD23. Experiments were repeated three times and the representative dotplot data were shown.
- Figure S5. The time course of binding and internalization of 3H-folic acid by thioglycollate-elicited macrophages (JPG, 47.2 KB)
-
- Figure S6. The effect of various cytokines/stimulants on the percentage of FR+ human monocytes-derived macrophages measured by flow cytometry (JPG, 82.4 KB)
-
The ratios of %FR+ macrophages under different cytokines/stimulants conditions over %FR+ macrophages in non-stimulated (control) condition are shown. The data is presented in average ± SD with n ≥ 3.
- Figure S7 Mouse bone marrow-derived macrophages activated in vitro do not express FR (JPG, 50.1 KB)
-
Mouse bone marrow-derived macrophages were primed with IFN-γ for 2 hours followed by LPS stimulation for 18 hours. Compared to non-stimulated counterparts (left panel), in vitro activated mouse bone marrow-derived macrophages (right panel) expressed a similar level of F4/80 but a higher level of CD86 and TNF-α. Neither non-stimulated nor IFN-γ/LPS stimulated mouse macrophages expressed FR.
|
|
