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

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Representative immunoblots showing CA nanotube mediated co-pelleting of endogenous CPSF6 (a), NUP153 (b) and SEC24C (c) from MT4 cell lysates. The proteins of interest were visualized by antibodies ab175237(Abcam) against CPSF6, NB100-93329 (Novus) against NUP153, ab122633 (Abcam) against SEC24C. Lane 1: cell lysate. Lane 2: supernatant or unbound fraction after pelleting in the absence of CA tubes. Lane 3: supernatant or unbound fraction after co-pelleting with pre-formed CA tubes. Lane 4: pelleted or bound fraction in the absence of CA tubes. Lane 5: pelleted or bound fraction in the presence of CA tubes. The experiments were repeated 3 times independently with similar results. Quantitation of GST-mediated affinity pull-down of native HIV-1 cores bound to indicated concentrations of GST-CPSF6_261-358(LCR-FG-LCR) vs GST-CPSF6(FG)/nonLCR (d), GST-NUP153_1306-1450(LCR-FG-LCR) vs GST-NUP153(FG)/nonLCR (e); and GST-SEC24C_196-314(LCR-FG-LCR) vs GST-SEC24C(FG)/nonLCR (f). The results were analyzed by Origin 2019 (v.9.6) software to determine binding K_d values. Each data point represents mean values + /− SD from three independent experiments. Source data are provided as a Source Data file. g Quantitation of GST-mediated affinity pull-down of isolated native HIV-1 cores vs crosslinked CA hexamers with 2 μM GST-CPSF6_261-358(LCR-FG-LCR) (left), GST-NUP153_1306-1450(LCR-FG-LCR) (middle) and GST-SEC24C_196-314(LCR-FG-LCR) (right). Mean values + /− SD from three independent experiments are shown. Source data are provided as a Source Data file. h Representative immunoblots of three independent experiments showing co-pelleting of GST-CPSF6_261-358(LCR-FG-LCR) (top), GST-NUP153_1306-1450(LCR-FG-LCR) (middle) and GST-SEC24C_196-314(LCR-FG-LCR) (bottom) with pre-formed CA nanotubes. The experiments were repeated 3 times independently with similar results.