g lisa kit  (Cytoskeleton Inc)

Journal: bioRxiv

Article Title: Pre-myelinating oligodendrocyte ADGRG1 is required for axon ensheathment and CNS myelin formation

doi: 10.64898/2026.01.26.701898

Figure Lengend Snippet: (A) Schematic of the proposed signaling pathway showing ADGRG1 activating the RhoA pathway to promote F-actin polymerization in preOLs. (B) Immunostaining of control and cKO preOLs for ADGRG1 (H11, green), GTP-RhoA (magenta), DAPI (blue) and brightfield. Colocalization between ADGRG1 and GTP-RhoA was observed in the processes of control preOLs, but both signals were downregulated in cKO preOLs. Scale bar, 10 μm. (C) Representative images of control and cKO oligodendrocyte cultures differentiated for 3, 5, or 7 days, treated with either vehicle, Rho inhibitor (for control), or Rho activator (for cKO). Cultures were stained for MBP (magenta), BCAS1 (green), Olig2 (gray), and DAPI (blue). Scale bar, 10 μm. (D) Quantification of the percentage of BCAS1 + Olig2 + cells in total Olig2 + cells across differentiation time points. Each circle represents one biological replicate. ** p=0.0011 (diff 3 day Con vs cKO), ** p=0.0011 (diff 3 day cKO vs cKO + Rho activator), ** p=0.0088 (diff 5 day Con vs Con + Rho inhibitor), * p=0.0103 (diff 5 day Con vs cKO), * p=0.0181 (diff 5 day cKO vs cKO + Rho activator), *** p=0.0005 (diff 7 day Con vs cKO), *** p=0.0003 (diff 7 day cKO vs cKO + Rho activator). Two-way ANOVA with Bonferroni’s multiple comparisons test. (E) Quantification of the percentage of MBP + Olig2 + cells in total Olig2 + cells across differentiation time points. Each circle represents one biological replicate. **** p<0.0001 (diff 5 day Con vs Con + Rho inhibitor), ** p=0.0047 (diff 5 day Con vs cKO), * p=0.0108 (diff 5 day cKO vs cKO + Rho activator), **** p<0.0001 (diff 7 day Con vs Con + Rho inhibitor), ** p=0.0010 (diff 7 day Con vs cKO), **** p<0.0001 (diff 7 day cKO vs cKO + Rho activator). Two-way ANOVA with Bonferroni’s multiple comparisons test. Data are presented as means ± SEM.

Article Snippet: The following compounds were used: RhoA Activator II (1 μg/mL, Cytoskeleton Inc., Cat. #: CN03-A) and RhoA Inhibitor I (1 μg/mL, Cytoskeleton Inc., Cat. #: CT04-A).

Techniques: Immunostaining, Control, Staining

Journal: bioRxiv

Article Title: Pre-myelinating oligodendrocyte ADGRG1 is required for axon ensheathment and CNS myelin formation

doi: 10.64898/2026.01.26.701898

Figure Lengend Snippet: (A) Schematic of the proposed signaling pathway showing ADGRG1 activating the RhoA pathway to promote F-actin polymerization in preOLs. (B) Immunostaining of control and cKO preOLs for ADGRG1 (H11, green), GTP-RhoA (magenta), DAPI (blue) and brightfield. Colocalization between ADGRG1 and GTP-RhoA was observed in the processes of control preOLs, but both signals were downregulated in cKO preOLs. Scale bar, 10 μm. (C) Representative images of control and cKO oligodendrocyte cultures differentiated for 3, 5, or 7 days, treated with either vehicle, Rho inhibitor (for control), or Rho activator (for cKO). Cultures were stained for MBP (magenta), BCAS1 (green), Olig2 (gray), and DAPI (blue). Scale bar, 10 μm. (D) Quantification of the percentage of BCAS1 + Olig2 + cells in total Olig2 + cells across differentiation time points. Each circle represents one biological replicate. ** p=0.0011 (diff 3 day Con vs cKO), ** p=0.0011 (diff 3 day cKO vs cKO + Rho activator), ** p=0.0088 (diff 5 day Con vs Con + Rho inhibitor), * p=0.0103 (diff 5 day Con vs cKO), * p=0.0181 (diff 5 day cKO vs cKO + Rho activator), *** p=0.0005 (diff 7 day Con vs cKO), *** p=0.0003 (diff 7 day cKO vs cKO + Rho activator). Two-way ANOVA with Bonferroni’s multiple comparisons test. (E) Quantification of the percentage of MBP + Olig2 + cells in total Olig2 + cells across differentiation time points. Each circle represents one biological replicate. **** p<0.0001 (diff 5 day Con vs Con + Rho inhibitor), ** p=0.0047 (diff 5 day Con vs cKO), * p=0.0108 (diff 5 day cKO vs cKO + Rho activator), **** p<0.0001 (diff 7 day Con vs Con + Rho inhibitor), ** p=0.0010 (diff 7 day Con vs cKO), **** p<0.0001 (diff 7 day cKO vs cKO + Rho activator). Two-way ANOVA with Bonferroni’s multiple comparisons test. Data are presented as means ± SEM.

Article Snippet: The following compounds were used: RhoA Activator II (1 μg/mL, Cytoskeleton Inc., Cat. #: CN03-A) and RhoA Inhibitor I (1 μg/mL, Cytoskeleton Inc., Cat. #: CT04-A).

Techniques: Immunostaining, Control, Staining

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