g lisa kit  (Cytoskeleton Inc)

Journal: bioRxiv

Article Title: Targeting AKAP13 RhoGEF activity ameliorates pro-fibrotic phenotypes driven by the IPF associated AKAP13 risk variant

doi: 10.64898/2026.01.21.700846

Figure Lengend Snippet: (A) RhoA activity measured by RhoA G-LISA in wild-type (blue) and rs62025270 (red) iHBECs treated with or without 50 µM LPA for 2 minutes. Data are presented as raw OD values at 490 nm (n = 3). (B) Real-time cell impedance analysis of wild-type (blue) and rs62025270 (red) iHBECs using the xCELLigence RTCA system. Data are shown as cell index over a 5-hour period (n = 3). (C) Representative immunoblot images of phospho-SMAD2 and total SMAD2/3 in iHBECs treated with or without 50 µM LPA for 2 hours. (D) Densitometric quantification of immunoblots shown in (C), presented as phospho-SMAD2 relative to total SMAD2/3 (n = 3). Statistical analysis was performed using a paired t-test between wild type and rs62025270 treated with LPA. * p<0.05, ** p<0.01.

Article Snippet: RhoA activity was measured using the RhoA G-LISA Activation Assay Biochem Kit (Cytoskeleton) following the manufacturer protocol.

Techniques: Activity Assay, Western Blot

Journal: bioRxiv

Article Title: Targeting AKAP13 RhoGEF activity ameliorates pro-fibrotic phenotypes driven by the IPF associated AKAP13 risk variant

doi: 10.64898/2026.01.21.700846

Figure Lengend Snippet: (A) Schematic representation of cDNA constructs encoding full-length AKAP13, Proto-Lbc, and △-Lbc. (B) RhoA activity measured by RhoA G-LISA in iHBECs transfected with empty vector, Proto-Lbc, or △-Lbc., followed by treatment with or without 50 µM LPA for 2 min. Data are presented as fold change relative to control (n = 3). (C) Representative immunoblot showing expression of FLAG-tagged Proto-Lbc and Δ-Lbc in iHBECs. (D) Real-time cell impedance measurement of iHBECs using the xCELLigence RTCA system. Data are shown as normalised cell index over a 5-hour period (n = 3).

Article Snippet: RhoA activity was measured using the RhoA G-LISA Activation Assay Biochem Kit (Cytoskeleton) following the manufacturer protocol.

Techniques: Construct, Activity Assay, Transfection, Plasmid Preparation, Control, Western Blot, Expressing

Journal: bioRxiv

Article Title: Targeting AKAP13 RhoGEF activity ameliorates pro-fibrotic phenotypes driven by the IPF associated AKAP13 risk variant

doi: 10.64898/2026.01.21.700846

Figure Lengend Snippet: (A) Real-time cell impedance analysis of wild-type (blue) and rs62025270 (red) iHBECs treated with 3 µM A13 or DMSO control using the xCELLigence RTCA system. Data are presented as cell index over a 5-hour period (n = 3). (B) RhoA activity measured by RhoA G-LISA in WT (blue) and rs62025270 (red) iHBECs treated with or without 50 µM LPA for 2 minutes. Data are presented as fold change relative to WT (n = 3). Statistical analysis was performed using the Friedman test for multiple comparisons within genotype. *p < 0.05 (C) Representative immunoblot images of phospho-SMAD2 and total SMAD2/3 in iHBECs treated with or without 50 µM LPA for 2 hours in the presence or absence of 10 µM A13. (D) Densitometric quantification of immunoblots shown in (C). Statistical analysis was performed using the Friedman test for multiple comparisons. *p < 0.05

Article Snippet: RhoA activity was measured using the RhoA G-LISA Activation Assay Biochem Kit (Cytoskeleton) following the manufacturer protocol.

Techniques: Control, Activity Assay, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: FGF12 induces aberrant mechanosignaling in aortic smooth muscle cells during thoracic aortic aneurysm formation in Marfan syndrome mice

doi: 10.1038/s12276-025-01621-y

Figure Lengend Snippet: a FGF12 induces RhoA activation in HASMCs. Levels of GTP-bound RhoA (RhoA-GTP) were measured by RhoA pulldown assays in untransduced and Ad-transduced HASMCs. b Losartan reduces FGF12 -induced RhoA activation. Following Ad transduction, HASMCs were treated with or without losartan. c FGF12 enhances phosphorylation of MLC (p-MLC) and cofilin (p-cofilin) via ROCK. Western blotting was performed on untransduced and Ad-transduced HASMCs treated with or without the ROCK inhibitor (Y27632). d FGF12 promotes nuclear translocation of MRTF-A and stress fiber formation via ROCK. Following Ad transduction, HASMCs were treated with or without Y27632 and stained for MRTF-A (green) and with phalloidin (red). e FGF12 upregulates integrin α5, integrin β1 and paxillin expression and enhances phosphorylation of SRC (p-SRC) and FAK (p-FAK) via ROCK. Western blotting was performed on untransduced and Ad-transduced HASMCs treated with or without Y27632. f FGF12 promotes FA formation via ROCK. Following Ad transduction, HASMCs were treated with or without Y27632 and stained for paxillin (green) and with phalloidin (red). g FGF12 increases cellular stiffness, which is suppressed by losartan or Y27632. After Ad transduction, HASMCs were treated with or without losartan and/or Y27632. The stiffness of individual cells was measured by AFM nanoindentation and is expressed as Young’s modulus ( n = 10–15 cells per group). In a – c and e , RhoA-GTP and phospho-protein levels were normalized to total protein. Protein levels are quantified relative to untransduced controls (Cont, set as 1; n = 3 independent experiments). In d and e , images and quantified data are representative of three independent experiments. Nuclei are stained with DAPI (blue). Scale bars, 20 μm. All data are presented as mean ± s.e.m. and were analyzed by one-way ANOVA with Bonferroni’s post-hoc test (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Active RhoA (GTP-bound RhoA) was measured using a RhoA activation assay kit (Cytoskeleton).

Techniques: Activation Assay, Transduction, Phospho-proteomics, Western Blot, Translocation Assay, Staining, Expressing