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. 2021 Apr 13;12:2198. doi: 10.1038/s41467-021-22522-4

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© The Author(s) 2021

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. 7 — a Left: expression of MRCKα and MRCKβ (myotonic dystrophy kinase-related Cdc42-binding kinase) genes in EFA6B KO55 cells analyzed by qPCR 2 days post-transfection with the indicated siRNAs. siMRCKα+β/α and siMRCKα+β/β indicate the gene expression level of MRCKα or MRCKβ after transfection with siRNAs against both MRCKs, N = 3 for single siRNA and N = 2 for combined siRNA. Middle: expression of pMLC and total MLC analyzed by immunoblot 2 days post-transfection of EFA6B KO55 cells with the indicated siRNAs. Right: quantification of the percentage of aggregates with invasive protrusions of EFA6B KO55 cells grown in collagen I for 2 days post-transfection with the indicated siRNAs. N = 3, average ±SEM, one-way ANOVA test with Dunnett’s multiple comparison p-values. b Expression of the indicated proteins analyzed by immunoblot two days post-transfection of EFA6B KO55 cells with the indicated siRNAs. ROCK: Rho-associated protein kinase. HSP60 served as a loading control. Right panel: quantification of the percentage of aggregates with invasive protrusions of EFA6B KO55 cells grown in collagen I for 2 days after transfection with the indicated siRNAs. N = 3, average ± SEM, one-way ANOVA test with Dunnett’s multiple comparison p-values. c Left: expression of N-WASP (Neural Wiskott–Aldrich Syndrome Protein) analyzed by immunoblot two days post-transfection of EFA6B KO55 cells with N-WASP targeted siRNAs. Actin served as a loading control. Right: quantification of the percentage of aggregates (n = 100) with invasive protrusions of EFA6B KO55 cells grown in collagen I for 2 days after transfection with N-WASP-directed siRNAs. N = 3, average ± SEM, one-way ANOVA test with Dunnett’s multiple comparison p-values. d Left: expression of ARP3 (Actin Related Protein 3) analyzed by immunoblot 2 days post-transfection of EFA6B KO55 cells with ARP3-targeted siRNAs. Actinin served as a loading control. Right: quantification of the percentage of aggregates (n = 100) with invasive protrusions of EFA6B KO55 cells grown in collagen I for 2 days after transfection with ARP3-targeted siRNAs. N = 3, average ± SEM, one-way ANOVA test with Dunnett’s multiple comparison p-values. Source data are provided as a Source Data file.