Journal: medRxiv

Article Title: A postzygotic GNA13 variant upregulates the RHOA/ROCK pathway and alters melanocyte function in a mosaic skin hypopigmentation syndrome

doi: 10.1101/2024.07.24.24310661

Figure Lengend Snippet: a,b Immunofluorescence images of Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP (green) expressed in NHEM. Left panels: The cells were labelled with phalloidin for visualising F-actin ( a ) or pMLC ( b ) (red), and Hoechst for nuclei (blue). Right panels: Quantifications of morphological parameters (perimeter, circularity and solidity) and F-actin content ( a ), and pMLC levels ( b ). c Quantification of RHOA-GTP levels in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP -expressing NHEM cells. Means +/- SEM are shown.

Article Snippet: Cells were incubated with primary antibodies anti-pMLC (Cell Signaling Technology), anti-vinculin (Life Technologies) or anti-RHOA-GTP (CurieCoreTech) for 1h at RT.

Techniques: Immunofluorescence, Expressing

Journal: medRxiv

Article Title: A postzygotic GNA13 variant upregulates the RHOA/ROCK pathway and alters melanocyte function in a mosaic skin hypopigmentation syndrome

doi: 10.1101/2024.07.24.24310661

Figure Lengend Snippet: a Quantification of RHOA-GTP levels in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP expressing B16-F0 cells that correspond to the mean intensity of the signal generated from anti-GTP RHOA antibody labelling. Quantification of F-actin content ( b ) and circularity ( c ) in RHOA WT-GFP and RHOA Q63L-GFP expressing B16-F0 cells. Quantification of F-actin content ( d ) and circularity ( e ) in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP expressing B16-F0 cells, upon RHOA inhibition treatment with CT04. Quantification of F-actin content ( f ) and circularity ( g ) in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP expressing B16-F0 cells, upon ROCK inhibition treatment with Y27632. Means +/- SEM are shown.

Article Snippet: Cells were incubated with primary antibodies anti-pMLC (Cell Signaling Technology), anti-vinculin (Life Technologies) or anti-RHOA-GTP (CurieCoreTech) for 1h at RT.

Techniques: Expressing, Generated, Inhibition

Journal: medRxiv

Article Title: A postzygotic GNA13 variant upregulates the RHOA/ROCK pathway and alters melanocyte function in a mosaic skin hypopigmentation syndrome

doi: 10.1101/2024.07.24.24310661

Figure Lengend Snippet: a,b Immunofluorescence images of Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP (green) expressed in NHEM. Left panels: The cells were labelled with phalloidin for visualising F-actin ( a ) or pMLC ( b ) (red), and Hoechst for nuclei (blue). Right panels: Quantifications of morphological parameters (perimeter, circularity and solidity) and F-actin content ( a ), and pMLC levels ( b ). c Quantification of RHOA-GTP levels in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP -expressing NHEM cells. Means +/- SEM are shown.

Article Snippet: Cells were incubated with primary antibodies anti-pMLC (Cell Signaling Technology), anti-vinculin (Life Technologies) or anti-RHOA-GTP (CurieCoreTech) for 1h at RT.

Techniques: Immunofluorescence, Expressing

Journal: medRxiv

Article Title: A postzygotic GNA13 variant upregulates the RHOA/ROCK pathway and alters melanocyte function in a mosaic skin hypopigmentation syndrome

doi: 10.1101/2024.07.24.24310661

Figure Lengend Snippet: a Quantification of RHOA-GTP levels in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP expressing B16-F0 cells that correspond to the mean intensity of the signal generated from anti-GTP RHOA antibody labelling. Quantification of F-actin content ( b ) and circularity ( c ) in RHOA WT-GFP and RHOA Q63L-GFP expressing B16-F0 cells. Quantification of F-actin content ( d ) and circularity ( e ) in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP expressing B16-F0 cells, upon RHOA inhibition treatment with CT04. Quantification of F-actin content ( f ) and circularity ( g ) in Gα 13 WT-YFP, Gα 13 R200K-YFP or Gα 13 Q226L-YFP expressing B16-F0 cells, upon ROCK inhibition treatment with Y27632. Means +/- SEM are shown.

Article Snippet: Cells were incubated with primary antibodies anti-pMLC (Cell Signaling Technology), anti-vinculin (Life Technologies) or anti-RHOA-GTP (CurieCoreTech) for 1h at RT.

Techniques: Expressing, Generated, Inhibition

Journal: iScience

Article Title: ADAMTS18-fibronectin interaction regulates the morphology of liver sinusoidal endothelial cells

doi: 10.1016/j.isci.2024.110273

Figure Lengend Snippet: Enhanced VEGFA and Rho signaling pathway in LSECs from Adamts18 −/− mice (A) Immunofluorescence staining for CD14 in primary LSECs from 2-week-old Adamts18 +/+ and Adamts18 −/− mice. Scale bar, 50 μm. (B) Representative western blots for VEGFA, pSrc, Src, RhoA-GTP, RhoA, pYAP, YAP, pMLC, and MLC in primary LSECs. (C) The relative quantity of VEGFA is normalized to that of GAPDH and phosphorylated proteins is normalized to that of corresponding total proteins. Data are expressed as mean ± SD ( n = 3). (D) Representative immunofluorescence staining for RhoA-GTP, pYAP, and pMLC in primary LSECs. Scale bar, 10 μm. (E) Quantification of the fluorescence intensity in panel D by ImageJ. Each LSEC slide was analyzed for 5 fields. Data are represented as mean ± SD ( n = 3). Each dot or square represents one individual. Ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01; two-tailed Student’s t test.

Article Snippet: Mouse Anti-RhoA-GTP Monoclonal Antibody , NewEast Biosciences , Cat# 26904, RRID: AB_1961799.

Techniques: Immunofluorescence, Staining, Western Blot, Fluorescence, Two Tailed Test

Journal: iScience

Article Title: ADAMTS18-fibronectin interaction regulates the morphology of liver sinusoidal endothelial cells

doi: 10.1016/j.isci.2024.110273

Figure Lengend Snippet:

Article Snippet: Mouse Anti-RhoA-GTP Monoclonal Antibody , NewEast Biosciences , Cat# 26904, RRID: AB_1961799.

Techniques: Recombinant, RNAscope, SYBR Green Assay, Modification, Staining, Peroxidation Assay, Immunoprecipitation, Activity Assay, Plasmid Preparation, Software