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. 2016 Nov 14;6:37021. doi: 10.1038/srep37021

Figure 2. Nef prevents MHC-I from entering into a Rab11 dependent recycling route.

Figure 2

(a) MHC-I-eGFP (panel 1) or Nef-VC and MHC-I-VN-Flag (panel 2) and dsRed-Rab11 were co-transfected into HeLa cells. 24 hrs post transfection, cells were fixed and mounted onto coverslips. GFP and BiFC fluorescence was observed under the FITC channel, and dsRed-Rab11a was observed under the Cy3 channel. (b) Co-localization was quantified by using the Pearson’s correlation through the JaCoP Plug-in on ImageJ. (c) MHC-I (BB7.2) uptake experiments were performed as described in the materials and methods. BiFC signal is visualized in green, and MHC-I uptake was pseudocolored in red, nuclei were counterstained in blue. (d) Percent of co-localization (left axis and black bars), and Pearson’s correlation (right axis and grey bars) were determined using the Mander’s and Pearson’s correlation respectively through the JaCoP Plug-in on ImageJ. Error bars were calculated by quantification of at least 50 cells between 3 independent experiments. (**Indicates p-value < 0.01, ***indicates p-value < 0.001).