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Fig. 3 | Retrovirology

Fig. 3

From: Mutagenesis of N-terminal residues of feline foamy virus Gag reveals entirely distinct functions during capsid formation, particle assembly, Gag processing and budding

Fig. 3

Intracellular capsid assembly of FFV Gag mutants determined by sedimentation and transmission electron microscopy analyses. a 293T cells were transfected in 10 cm dishes with 8 µg of proviral pCF-7-based Gag mutants (mLNPLQ, mLQQLY, mINNGL, mQPNPG, mHGDII, mAVRFT, mGGPWG or mPGDRW) or the parental provirus pCF-7. Two days p.t., cytoplasmic extracts were prepared and used for sucrose gradient sedimentation. Each six fractions (S1S6) were collected from the top of the gradient and analyzed by immunoblotting. Positions of FFV Gag p48 and p52 are marked. b Intracellular capsid assembly of wt Gag and Gag mutants mLQQLY and mHGDII were visualized by transmission electron microscopy of transfected and fixed 293T cells. Transmission electron micrographs show representative thin sections of 293T cells transiently transfected with pCF-7-based plasmids expressing mLQQLY, mHGDII or wt Gag. Capsids are marked by black arrows. Scale bars are 500 nm in length

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