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. 1998 Oct 5;188(7):1359–1368. doi: 10.1084/jem.188.7.1359

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Protein and mRNA expression of α_vβ₅ and CD36 are downregulated during DC maturation. Immature DCs (A) and mature DCs (B) were incubated with anti-α_vβ₃ (clone 23C6; PharMingen), anti-CD36 (clone FA6; obtained from the fifth international workshop on leukocyte differentiation antigens), or anti-α_vβ₅ (clone P1F6; Chemicon International, Inc.), followed by PE-conjugated GAM-Ig (Sigma Biosciences). All cells were analyzed by FACScan^®. (C) RNA was isolated from highly purified sorted cell populations of immature and mature DCs as previously described. Reverse transcription was carried out and after 30 cycles of PCR the distinct bands for β₃, β₅, and CD36 could be seen in the immature DCs (lane 1). In mature DCs, only a faint band for CD36 and no band for β₅ could be visualized (lane 2), indicating minimal mRNA. In contrast, a band was evident for β₃ in the mature DCs. Note that the doublet for β₃ is an artifact in this particular exposure and does not indicate two unique bands (lane 2). As a positive control, extracts from Bowes melanoma cells, which are known to express β₃ and CD36 (29), were run (lane 3). As negative control, the Bowes melanoma cells were run in the absence of a reverse transcriptase (lane 4).

Protein and mRNA expression of α_vβ₅ and CD36 are downregulated during DC maturation. Immature DCs (A) and mature DCs (B) were incubated with anti-α_vβ₃ (clone 23C6; PharMingen), anti-CD36 (clone FA6; obtained from the fifth international workshop on leukocyte differentiation antigens), or anti-α_vβ₅ (clone P1F6; Chemicon International, Inc.), followed by PE-conjugated GAM-Ig (Sigma Biosciences). All cells were analyzed by FACScan^®. (C) RNA was isolated from highly purified sorted cell populations of immature and mature DCs as previously described. Reverse transcription was carried out and after 30 cycles of PCR the distinct bands for β₃, β₅, and CD36 could be seen in the immature DCs (lane 1). In mature DCs, only a faint band for CD36 and no band for β₅ could be visualized (lane 2), indicating minimal mRNA. In contrast, a band was evident for β₃ in the mature DCs. Note that the doublet for β₃ is an artifact in this particular exposure and does not indicate two unique bands (lane 2). As a positive control, extracts from Bowes melanoma cells, which are known to express β₃ and CD36 (29), were run (lane 3). As negative control, the Bowes melanoma cells were run in the absence of a reverse transcriptase (lane 4).