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Fig. 3 | Cell Regeneration

Fig. 3

From: RhoA/Rock activation represents a new mechanism for inactivating Wnt/β-catenin signaling in the aging-associated bone loss

Fig. 3

RhoA/Rock activates Jak1/2 and Gsk3β to destabilize β-catenin. a-c Western analyses in C3H10T1/2 cells transfected with or without RhoA-si or Rock1 + Rock2 siRNA (Rock1,2-si) and treated with or without rWnt3a at 100 ng/ml for the indicated time or 1 h. d, e Western or Lef1-luciferase expression analyses in C3H10T1/2 cells transfected with Gsk3β variants and treated with rWnt3a for 6 or 48 h, respectively. f-k Western analyses in C3H10T1/2 cells transfected with RhoA-si, caRhoA, caRock2, caJak1/2, infected with lentiviral Jak2-shRNA (Jak2-sh), or treated with P6 at 50 nM, followed by incubation with rWnt3a for the indicated times or 1 h. l Co-immunoprecipitation by using IgG1 or Gsk3β antibody in 293 cells transfected with HA-Jak1/2 and Myc-Gsk3β. m In vitro phosphorylation of GSK3β protein by active JAK2 in kinase assay buffer with or without ATP. Phosphorylated proteins were normalized to their total amounts, respectively. Mean ± SD, *, ¶ p < 0.05, **, ++p < 0.01, n = 4, Tukey-Kramer multiple comparisons test

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