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. 2021 May 11;17(5):e1009580. doi: 10.1371/journal.ppat.1009580

Fig 7. Dynein light chain DYNLT3 facilitates nuclear delivery of incoming HPV16.

Fig 7

(A) RNAi of various dynein light chains by each two individual siRNAs as indicated in HeLa cells was followed by HPV16-PsV infection. Infectivity was scored based on cells expressing GFP over total cells. The infectivity was normalized to control siRNA transfected cells as relative (rel.) infection. Error bars indicate the SD of three independent experiments. (B)(D) A L2 chromosomal association assay after DYNLT3 (B) or DYNLL2 (D) depletion in HeLa Kyoto_H2B-mCherry_L2-GFP cells was performed as in Fig 2B. (C)(E) CAI of panels (B)(D). At least 40 cells were analyzed in three independent experiments, and the median was indicated by the black bar. (F) Caveolin1-HA, L2-3xHA or HA-RanBP10 were expressed together with FLAG-DYNLT3 in HEK293 cells. Immunoprecipitation against HA was performed on cell lysates. Caveolin1-HA was used as a negative control of immunoprecipitation. HA, FLAG, and GAPDH were detected by Western Blotting after immunoprecipitation. (G) Knockdown of DYNLT3 in HeLa cells was followed by infection with EdU-labelled HPV16. Cells were fixed at 20 h.p.i., and stained with an anti-P230 antibody and Hoechst-33258 to indicate the TGN and nucleus, respectively. (H) Quantification of co-localized vDNA signals with nucleus or P230. At least 35 cells were analyzed in three independent experiments. The error bars indicate the SD.