Figure 2 : Decrease in fluorescence intensity and increase in FRET ratio in the nucleus of HeLaP4, C8166 T and primary CD4+ T cells.

From: Dynamic Oligomerization of Integrase Orchestrates HIV Nuclear Entry

Figure 2

(A) Representative confocal image of a lamin immunostained (blue) HeLaP4 cell infected with VSV-G pseudotyped HIVIN-eGFP for 6 h. Red arrows point towards nuclear IN-eGFP complexes. (B) Fluorescence intensity of HIVIN-eGFP complexes in infected HeLaP4 cells in the cytoplasm (black circle) and in the nucleus (grey circle), obtained using the automated image analysis tool. Box-plot whiskers represent 5th and 95th percentile, the median value is represented by the line within the box, and the square box depicts the mean. (C) Mean FRET ratios of HIVIN-mTFP1+IN-mVenus complexes in infected HeLaP4 cells in the cytoplasm (white) and in the nucleus (grey). (D) The left panel shows a representative confocal image of lamin immunostained (blue) C8166 T cells infected with HIVEnvIN-eGFP, containing a wild-type HIV envelope, for 24 h. Red arrows indicate nuclear IN-eGFP complexes. Fluorescence intensity (middle) and mean FRET ratios (right) of HIV complexes in C8166 T cells infected with HIVEnvIN-eGFP and HIVEnvIN-mTFP1+IN-mVenus, respectively. (E) The left panel shows a representative confocal image of DAPI stained (blue) CD4+ T cells infected with HIVEnvIN-eGFP, containing a wild-type HIV envelope, for 24 h. Red arrows indicate nuclear IN-eGFP complexes. Fluorescence intensity (middle) and mean FRET ratios (right) are shown for HIV complexes in primary CD4+ T cells infected with HIVEnvIN-eGFP and HIVEnvIN-mTFP1+IN-mVenus, respectively. Error bars represent standard error of mean (SEM); ***p-value < 0.001, **p-value < 0.01 obtained with an unpaired Two Sample t-test with unequal variance of the data. Scale bar represents 5 μm.