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. 2011 Dec;85(23):12482–12491. doi: 10.1128/JVI.05218-11

Fig. 1.

Fig. 1.

Detection of the feline elonginB, elonginC, and cullin5 gene transcripts and the fElonginC and fCul5 proteins in CrFK cells. (A) PCR amplification of elonginB, elonginC, and cullin5 cDNA by RT-PCR from total mRNA of feline CrFK cells. The β-actin cDNA was used as a control for the RNA quality of each sample. Mk indicates the DNA molecular marker, while control is the mock DNA-negative control. (B) Cell lysates of 293T and CrFK cells were analyzed by Western blotting using anti-hCul5, anti-hElonginC, and anti-hElonginB antibodies. The control is a loading control. (C) 293T cells (106) were transfected with 1 μg of VR-hCul5-myc, VR-hEB-HA, VR-hEC-HA, VR-fCul5-myc, VR-fEB-flag, or VR-fEC-HA. Cells were harvested 48 h after transfection and analyzed by Western blotting with anti-HA, anti-myc and anti-flag antibodies. The control is nontransfected cells. (D) Alignment of the amino acid sequences of the feline ElonginB and human ElonginB. (E) Alignment of the amino acid sequences of the feline ElonginC and human ElonginC. (F) Alignment of the partial amino acid sequences of the feline Cul5 and human Cul5. The arrow indicates the single amino acid difference.