Fig. 4: PLXNB2 interacts with SEMA4C to promote homotypic clustering of breast tumor cells. | Nature Communications

Fig. 4: PLXNB2 interacts with SEMA4C to promote homotypic clustering of breast tumor cells.

From: Computational ranking identifies Plexin-B2 in circulating tumor cell clustering with monocytes in breast cancer metastasis

Fig. 4: PLXNB2 interacts with SEMA4C to promote homotypic clustering of breast tumor cells.The alternative text for this image may have been generated using AI.

a Schematic of PLXNB2-SEMA4C intercellular interactions in homotypic tumor cell clustering between two cancer cells. b Immunoblots of PLXNB2 (PB2) and SEMA4C expression in different breast cancer cell lines and PDX models using antibodies that are specific to detect both human and mouse isoforms. c Immunoblots of SEMA4C, CDC42, and PLXNB2 in the protein complex immunoprecipitated by anti-PLXNB2 antibody compared to IgG. d Immunoblot images showing reduced SEMA4C levels after SEMA4C KD in MDA-MB-231 tumor cells (left) and depleted PLXNB2 in PLXNB2 KO1 and KO2 cells (right). e Schematic illustration and immunoblotting of MDA-MB-231 control and PLXNB2-KO cells transfected with siRNA control or siSEMA4C knockdown, followed by tumor cell clustering assessment in suspension. f, g Representative images (f) and average cluster area/size curves (g) of MDA-MB-231 control (Con) and PB2 KO (KO) cells transfected with control siRNA (siCon) or SEMA4C siRNAs (si4C) for KD, measured by IncuCyte, n = 5 technical replicates examined over 5 independent experiments. For cluster size (area) comparisons with the group of Con+siCon, Student’s t test p = 0.033 for Con+si4C, p = 0.021 for KO+si4C, and p = 0.016 for KO+siCon. For the cluster number comparisons with the Control group, p < 0.001 for all three pairs above. P value were calculated using one-sided ANOVA. h Principal component analysis clusters of MDA-MB-231 single cells, siCon clusters, and siPLXNB2 clusters analyzed from global proteomics analysis with n = 3 technical replicates/ group over three independent experiments. i Heat map of differentially expressed proteins from global proteomics analysis of siCon single cells (blue), siCon clusters (green), and siPLXNB2 clusters (red), with n = 3 technical replicates/ group over three independent experiments. j Gene ontology biological processes analysis of significantly up-regulated and down-regulated proteins in PLXNB2 control vs. PLXNB2 KD clusters in breast cancer cells analyzed by mass spectrometry. Data are presented as mean values ± SD. P values were calculated using one-sided ANOVA. Source data are provided as a Source Data file.

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