Fig 1: Abl kinases Stabilize Ptpn12 to Regulate p130Cas Phosphorylation.(A) Deletion of Abl/Arg in MEF cells led to an increased phosphorylation of p130Cas. (B) Elevated expression of phosphorylated p130Cas was observed in Abl/Arg mutant lenses (arrowheads). (C) Inhibition of Abl kinase activity in NIH3T3 cells by Imatinib led to increased phosphorylation of p130Cas and enhanced Crk-p130Cas interaction, as demonstrated by co-immunoprecipitation. (D) Analysis from the DepMap database indicates positive co-dependency between Ptpn12 and Abl and negative co-dependency with Crk and p130Cas. (E) Expression of constitutively active AblPP in HEK293 cells led to tyrosine phosphorylation of Ptpn12 and an increase in its expression level. (F) Mutating of tyrosine residues in Ptpn12 significantly reduced its expression, which was mitigated by proteosome inhibitor MG132. (G) Increased p-p130Cas levels were observed following siRNA-mediated knockdown of Ptpn12, but not Ptpn14. (H) Deletion of p130Cas in Abl/Arg mutants rescued the lens vesicle separation phenotype. (I) Quantification of the lens stalk phenotype.
Fig 2: FGF and Abl signaling regulates lens vesicle separation via the p130Cas-Crk-Rac1-RhoA axis.(A) Cdh1 deletion did not exacerbate the lens vesicle separation defect in Abl/Arg mutants. (B) Combined ablation of Cdh2 and 3 did not disrupt lens development. (C) Phosphorylated MLC (pMLC) was downregulated in Abl/Arg mutant lens vesicles (arrowheads). (D) The corneal-lens attachment phenotype was similar in Abl/Arg and RhoALSL-G17V single mutants, but was significantly more severe in the double mutants (arrowheads). (E) Quantification of the corneal-lens contact length. One-way ANOVA n=3 for Abl/Arg and RhoALSL-G17V single mutants, n=5 for double mutants. (F) Rac1 deletion rescued lens vesicle separation in RhoALSL-G17V, Abl/Arg, KrasG12D and Tg-Fgf3 mutants. (G) Quantification of the lens stalk phenotype. (H) The model FGF and Abl kinase signaling in lens vesicle development. FGF and Abl kinase signaling regulates the p130Cas via Src phosphorylation and Ptpn12 dephosphorylation, respectively. This modulation affects the recruitment of Crk and CrkL to p130Cas and the activation of Erk and Rac1, the latter antagonizes RhoA activity. RhoA regulates actomyosin contractility, crucial for separating the lens vesicle from the surface ectoderm.
Supplier Page from Sino Biological, Inc. for Human PTPN12 Gene ORF cDNA clone expression plasmid, N-HA tag