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. 2021 Feb 28;13(3):386. doi: 10.3390/v13030386

Figure 3.

Figure 3

(a–c) Viruses spun on glass, fixed, and stained with AG3.0 antibody were produced with a non-fluorescent construct (pNL4-3-D116N-dEnv) that allows particle maturation by coding for HIV-1 Pol and high signal level in low excitation conditions. Green (mNeonGreen, sfGFP, or mWasabi), Red (mRuby3), Blue (mAB AG3.0 and anti-mouse-AF647) (d–f) As non-fluorescent particles are possible with trans complementation to pNL4-3 because the molecular clone does not code for any fluorescent protein, we quantified the frequency of labeled particles. We measured high levels of colocalization between mature particles, as identified by AG3.0 antibody, and the tagged IN, ranging from 75% to 90% colocalization. Colocalization was defined by centroids within 500nm of each other. Data represents one of the three repeated experiments with multiple technical repeats (>20) per experiment and thousands of viral particles analyzed (as shown in b).