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Fig. 5 | BMC Microbiology

Fig. 5

From: Structure-function analysis of Avian β-defensin-6 and β-defensin-12: role of charge and disulfide bridges

Fig. 5

Expression of chicken chemokine receptors CCR2 and CCR6 in CHO-K1 cells. a Fluorescence images of CCR2-GFP, CCR6-GFP and GFP proteins (green) expressing CHO-K1 cells. Transfected cells were fixed with methanol: acetone (v:v, 1:1), and nuclei were stained with 0.1 μg/ml DAPI (blue). b Gel electrophoresis of RT-PCR products of CCR2 (1,065 bp) and CCR6 (1,089 bp) derived from transfected CHO-K1 cells. GAPDH (125 bp) was included as RT-PCR control. c Western-blotting analysis of the CCR2/CCR6-GFP fusion protein expressed in CHO-K1 cells. Wild-type and only GFP expressing CHO-K1 cells were included as negative and positive controls, respectively. The protein fractions were probed with polyclonal goat anti-GFP primary antibody (1:1000) and HRP-coupled anti-goat secondary antibody (1:500), and visualized after color development with 4-CN and 30 % hydrogen peroxide. GFP-CCR2 is primarily located in the cytoplasmic membrane, GFP-CCR6 in nuclear membrane, and GFP alone in cytoplasm. Scale bar: 100 μm

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