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Fig. 3 | Retrovirology

Fig. 3

From: HIV-1 Vpu utilizes both cullin-RING ligase (CRL) dependent and independent mechanisms to downmodulate host proteins

Fig. 3

MLN4924 alleviates Vpu- but not Nef-mediated degradation of CD4. a Cultured TCM were infected as described in Fig. 2a. To assess total levels of CD4, cells were permeabilized and stained 24 h after addition of MLN4924 and analyzed by flow cytometry. Histograms are color-coded as described in Fig. 2a. b MFI values of total (intracellular) CD4 expression levels from DHIVGFP(Vpu+Nef−). Data was normalized and statistical significance obtained as described in Fig. 2b. n.s. not significant. c Primary CD4+ T cells were either mock infected or infected at an MOI of 1 with DHIV WT, DHIV Vpu−/Nef+ or DHIV Vpu+/Nef−. 2 days post infection, cell cultures were treated with either DMSO or 500 nM MLN4924. 24 h post MLN4924 treatment, cells were assessed for surface levels of CD4 between p24Gagneg (blue line) and p24Gagpos (red line). Gray shaded histograms represent an IgG matched isotype control. Shown is one representative experiment out of three. d Relative CD4 surface levels were quantified from data obtained in Fig. 3c and are depicted graphically as ±SEM of either cells treated with DMSO (left) or 500 nM MLN4924 (right). Statistical significance between p24Gagneg and p24Gagpos populations was determined as in Fig. 2b.

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