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. 2022 Oct 6;13:5879. doi: 10.1038/s41467-022-33662-6

Fig. 3. CPSF6 LCR effects on HIV-1 nuclear import and targeting to NS.

Fig. 3

a–c HEK293T CKO cells were back-complemented with indicated HA tagged CPSF6 (CPSF6) proteins and infected with VSV-G pseudotyped HIV-1 viruses (MOI 1) labeled with INsfGFP (green) for 4 h. Cells were fixed and immunostained for SON nuclear speckle (NS) marker (red) and nuclei with SiR-Hoechst (blue). a Single Z-stack images. Scale bar is 5 µm. b Quantitation of the number of IN-labeled viral replication complexes (VRCs) >0.5 µm of the nuclear periphery. c Quantification of IN-VRCs colocalized with SON-NSs. Mean values obtained from each experiment and SEM from a total of 4 independent experiments are shown in b, c. Each independent experiment analyzed >30 nuclei for each construct. Statistical significances of comparison of CPSF6/WT versus CPSF6/AD, CPSF6/NE, CPSF6/FU, CPSF6/CD and CKO (in b, c) were determined by Student’s two-sample, two-tailed t-test. P-values of CPSF6/AD, CPSF6/NE, CPSF6/FU, CPSF6/CD and CKO are 1.1E-05, 1.7E-05, 0.6, 0.09, 1.7E-05 (in b) and 8.8E-05, 9.2E-05, 0.69, 0.22, 8.5E-05 (in c), respectively. P > 0.05 was considered not significant (ns) and p < 0.001 was considered highly significant (***). Source data are provided as a Source Data file. CPSF6/AD CPSF6/ADD2, CPSF6/NE CPSF6/NEURM, CPSF6/FU CPSF6/FUS, CPSF6/CD CPSF6/CDK19, CPSF6/WT CPSF6/wild-type CPSF6, CKO CPSF6 knock-out, LCR low complexity region.