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Fig. 6 | Retrovirology

Fig. 6

From: Moloney leukemia virus 10 (MOV10) inhibits the degradation of APOBEC3G through interference with the Vif-mediated ubiquitin–proteasome pathway

Fig. 6

MOV10 binds with ElonginC or Cullin 5. a The knockdown efficiency of siElonginB, siElonginC and siCullin 5. 293T cells were transfected with siElonginB, siElonginC or siCullin 5, after 48 h, the cells were collected and detected with qRT-PCR. Data in A represents mean ± SD (error bars). b–i Co-immunoprecipitated analysis of the interaction between MOV10 and ElonginB (b), ElonginC (c, f, h), Cullin 5 (d, g, i), or CBF-β (e). pcDNA3.1-ElonginB-FLAG plus siElonginC and siCullin 5 (b), pcDNA3.1-ElonginC-FLAG plus siElonginB and siCullin 5 (c, h), pcDNA3.1-Cullin 5-FLAG plus siElonginB and siEloingC (d, i) or pcDNA3.1-CBF-β-FLAG (e) was transfected into 293T cells with pcDNA3.1-MOV10-HA or pcDNA3.1-GFP-HA. 293T cells were transfected with pcDNA3.1-MOV10-HA (pcDNA3.1-GFP-HA as a control) plus siElonginB and siCullin 5 (f) or siElonginB and siEloingC (g). After 48 h, the cells were collected and immunoprecipitated with anti-HA agarose beads (b–i). The samples in h and i were treated with RNase mixture. And then, immunoprecipitated samples were analyzed by immunoblotting with anti-FLAG, anti-HA, anti-GAPDH, anti-MOV10, anti-ElonginC, and anti-Cullin 5 antibodies. Empty vector pcDNA3.1 was used to equalize DNA amounts in each transfection. Values in h and i represent portions of ElonginC-FLAG/Cullin 5-FLAG normalized against MOV10-HA and compared with control. Data in a–i is representative of at least three independent experiments

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Retrovirology

ISSN: 1742-4690

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