Figure 1. RhoGAP activity measured as GTP hydrolysis by RhoA protein. Each reaction contained reaction buffer + GTP with the addition of RhoA alone (RhoA), RhoGAP alone (GAP), or RhoA + RhoGAP (RhoA + GAP). Reactions were incubated at 37°C for 20 min. Phosphate generated by hydrolysis of GTP was measured by the addition of CytoPhos™ reagent and reading of absorbance at 650 nm.
Rho activation in Swiss 3T3 fibroblasts. F-actin is visualized with fluorescent green phalloidin staining (Cat.# PHDG1). DAPI is the blue nuclear stain. Phalloidin staining shows F-actin-rich stress fibers. Cells were activated with Cat.# CN04.

Recently, Zhan et al. investigated the molecular pathways underlying human epidermal stem cell (hESC) migration during wound repair to better understand the activity of these cells in the restoration of skin cell and hair follicle homeostasis during wound repair and healing. The authors discovered that nitric oxide (NO) stimulates hESC migration during wound repair and healing, and using the NO donor S-nitroso-N-acetylpenicillamine (SNAP), demonstrated that NO promotes the migration of hESCs in vivo and in vitro. Furthermore, NO-mediated hESC migration in vitro requires cGMP-mediated activation of RhoA and Rac1 (but not Cdc42) signaling pathways. These signaling pathways were examined given that cell migration requires dynamic cell morphology changes which necessitate the re-arrangement of a cell’s actin cytoskeleton, which is strongly regulated by Rho-family GTPases. Cytoskeleton’s Cdc42, Rac1, and RhoA pull-down activation assays (Cat. # BK034, BK035, BK036, respectively) were essential in this study, providing researchers with the necessary research tools to measure Rho-family GTPase activities in a sensitive and consistent manner. These results provide valuable insight into the essential role that Rho-family GTPases have in NO-mediated hESC migration during wound repair and healing to restore proper cellular homeostasis.

Link to citation: 

Zhan R. et al. 2016. Nitric oxide promotes epidermal stem cell migration via cGMP-Rho GTPase signaling. Sci. Rep. 6, 30687.

Products used in this citation:

Cdc42 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 50 Assays, Cat. # BK034

Rac1 Pull-down Activation Assay Biochem Kit (bead pull-down format) - 50 Assays, Cat. # BK035

RhoA Pull-down Activation Assay Biochem Kit (bead pull-down format) - 80 Assays, Cat. # BK036

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alex castellanos