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. Author manuscript; available in PMC: 2015 May 20.
Published in final edited form as: Circulation. 2014 Apr 28;130(2):180–191. doi: 10.1161/CIRCULATIONAHA.113.005991

Figure 4.

Figure 4

ERK5 regulates phagocytic capacity of macrophages via Mfg-e8 secretion. A and B, Reduced phagocytic capacity of ERK5-MKO macrophages was restored by serum from nontransgenic littermate control (NLC) mice (A) or purified recombinant Mfg-e8 (B). NLC and ERK5-MKO bone marrow–derived macrophages were pretreated with serum from NLC or ERK5-MKO mice (A) or purified Mfg-e8 (11 μg/mL; B) for 24 hours, and the phagocytic index was analyzed by incubation with 5-chloromethylfuorescein diacetate–labeled apoptotic cells at a 1:5 (bone marrow–derived macrophages:apoptotic cells) ratio for 60 minutes, as described in Figure 1. Data are mean±SD (n=5); *P<0.05, **P<0.01. C, Decreased levels of Mfg-e8 in the serum of ERK5-MKO mice as quantified by enzyme-linked immunosorbent assay. Data are mean±SD (n=4); **P<0.01. D, Circulating Mfg-e8 was determined by immunoprecipitating (IP) Mfg-e8 from serum followed by immunoblotting (IB) with anti-Mfg-e8 (top); graph represents densitometry data (bottom). Data are mean±SD (n=4); **P<0.01. E, ERK5 regulates opsonin expression in the liver. Relative mRNA levels of indicated opsonins in the liver of NLC and ERK5-MKO mice were assessed by quantitative reverse transcription polymerase chain reaction. Data are mean±SD (n=5); *P<0.05, **P<0.01. Comparison between 2 independent groups was subjected to Wilcoxon rank sum test.