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. 2023 Jun 8;14(6):351. doi: 10.1038/s41419-023-05869-y

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© The Author(s) 2023

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. 4 — A Silver staining of C3b-binding nuclear proteins in A549-PTX cells prepared by Co-IP. B The top 20 enriched signaling pathways based on the 195 C3-binding nuclear proteins in A549-PTX cells. Verification of the interaction between C3b and RBBP4/7 by mutual Co-IP assays with anti-C3/C3b antibody (C) or anti-RBBP4/7 antibodies (D) or by immunocytochemistry assay (E) in A549-PTX cells. Scale bar, 25 μm. F C3b interacted with RBBP4/7-containing SIN3A complex but not NuRD or PRC2 complex in A549-PTX cells determined by Co-IP assay using anti-C3/C3b antibody followed by immunoblotting assay using the indicated antibodies. The SIN3A complex contains SIN3A, HDAC1/2, RBBP4/7, SDS, ING2, SAP18 and SAP30 subunits, while the NuRD complex contains MTA1 and CHD4 subunits, and the PRC2 complex contains EZH2 and SUZ12 subunits. G The interaction of C3b with the SIN3A complex was further confirmed by Co-IP and immunoblotting assays in A549-ORI cells with ectopic C3 expression. H A GST pull-down assay validated the interaction of C3b with the subunits of the SIN3A complex. I Identification of the physiological interaction of C3b and the SIN3A complex in A549-PTX cells by gel chromatography and immunoblotting. C3b assembles a new complex with the SIN3A complex containing SIN3A, HDAC1/2, RBBP4/7, ING2 and SAP18/30. The fraction numbers and elution of molecular weight markers with arrows are indicated.