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. 2023 Feb 1;222(3):e202206078. doi: 10.1083/jcb.202206078

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© 2023 Xue et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S2. — Y118-Paxillin exhibits distinct phosphorylation status in migrating cancer cells in vivo versus in vitro. (A) Top: pY118-Paxillin immunostaining (magenta) of ZMEL-GFP (GFP immunostaining, green) plated on in vitro cell culture dishes. Middle: pY118-Paxillin immunostaining (magenta) of ZMEL-mCherry (mCherry immunostaining, pseudo-colored green) in larval zebrafish (3 d post-transplantation). Bottom: pY118-Paxillin immunostaining (magenta) of the zebrafish developing heart (5 dpf). (B) Western blot showing the specificity of the pY118-Paxillin antibody and that it does not recognize Y118E-Paxillin and Y118F-Paxillin. (C) Representative images of ZMEL-GFP cells plated on 2D surfaces of different stiffnesses (left) and stained for pY118-Paxillin (right). (D) Unmodified Western blot of panels shown in Fig. 3 C—YUMM1.7 cells plated in culture and YUMM1.7 melanoma tumors in vivo blotted with Paxillin and pY118-Paxillin antibodies. “P” is parental cell line with no GFP expression. GFP was used as the loading control and as a control for the number of YUMM1.7 cells in mouse tumors. Source data are available for this figure: SourceData FS2.