Journal: JACC: Basic to Translational Science

Article Title: Ubiquitin Specific Protease 9X Regulates the Activation of ARK5 and Promotes Progression of Fibrotic Remodeling

doi: 10.1016/j.jacbts.2025.02.014

Figure Lengend Snippet: ARK5 Facilitates TGF-β1–Induced Myofibroblast Transformation via Activating RhoA/MRTF-A Pathway (A) Fibroblasts were infected with USP9X siRNA, then subjected to TGF-β1 stimulation. Total RNA samples were collected and sequenced. The most significantly altered KEGG pathways were displayed. (B) RhoA activity was assessed using western blot analysis after transfecting CFs with ARK5 siRNA or siRNA-control. (C) CFs were infected with siRNA-control, siRNA-USP9X, Ad-GFP, or Ad-ARK5, and then tested for RhoA activity. (D) MRTF-A protein levels in the nuclear and cytosolic fractions were detected by western blot analysis. (E) Immunofluorescence staining was performed to detect the intensity of MRTF-A. Scale bar: 50 μm. (F) CFs were infected with siRNA-control, siRNA-USP9X, Ad-GFP, or Ad-ARK5, and then tested for MRTF-A protein levels. (G) Representative immunofluorescence staining of MRTF-A for each group. Scale bar: 50 μm. n = 3-5, Values represent the mean ± SEM. One-way analysis of variance followed by Tukey’s post hoc test was used for statistical analysis. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Abbreviations as in , , and .

Article Snippet: Antibodies against USP9X (55054-1-AP), α-SMA (14395-1-AP), mechanosensitive transcription factor myocardin-related transcription factor-A (MRTF-A) (21166-1-AP), Ubiquitin (10201-2-AP), glyceraldehyde 3-phosphate dehydrogenase (60004-1-Ig), PCNA (10205-2-AP), and RhoA (10749-1-AP) were purchased from Proteintech.

Techniques: Transformation Assay, Infection, Activity Assay, Western Blot, Control, Immunofluorescence, Staining

Journal: Annals of surgical oncology

Article Title: Increased RhoA Activity Predicts Worse Overall Survival in Patients Undergoing Surgical Resection for Lauren Diffuse-Type Gastric Adenocarcinoma.

doi: 10.1245/s10434-016-5357-2

Figure Lengend Snippet: FIG. 1 a Schematic overview of the RhoA signaling pathway. RhoA and its downstream effector Rho-associated protein kinase (ROCK) have key regulatory roles in cell cycle regulation, actin-myosin- dependent cell contractility, cell motility, and cell proliferation. RhoA cycles between a GDP-bound inactive form and a GTP-bound active form. This process is mediated by guanine nucleotide exchange factor (GEF) and GTPase activating protein (GAP). Protein kinase A (PKA) phosphorylation of active Rho-GTP induces a tight association with

Article Snippet: Silencing of RhoA was achieved via lentiviral transduction of human RhoA short hairpin (sh) RNA (sc-29471-V; Santa Cruz Biotechnology).

Techniques: Phospho-proteomics

Journal: PLoS ONE

Article Title: MiR-124 Promote Neurogenic Transdifferentiation of Adipose Derived Mesenchymal Stromal Cells Partly through RhoA/ROCK1, but Not ROCK2 Signaling Pathway

doi: 10.1371/journal.pone.0146646

Figure Lengend Snippet: (A) QRT-PCR analysis of miR-124 expression in ADMSCs infected with the pLV-miR-124 locker lentiviral particles or the vector negative control (Vector NC). (B) QRT-PCR analysis of NSE, Tuj-1 and GFAP mRNA expression in ADMSCs (with or without miR-124 knockdown) on day 1 post-induction. (C) Representative images of flow cytometry analysis of ADMSCs (with or without miR-124 knockdown) with positive neurogenic markers on day 1 post-induction. (D) Quantification of the proportion of ADMSCs with positive neurogenic markers showed in (C). * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: The ready-to-use human RhoA shRNA lentiviral particles (sc-44209-V), ROCK1 shRNA lentiviral particles (sc-29473-V) and ROCK2 shRNA lentiviral particles (sc-29474-V) were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Expressing, Infection, Plasmid Preparation, Negative Control, Knockdown, Flow Cytometry

Journal: PLoS ONE

Article Title: MiR-124 Promote Neurogenic Transdifferentiation of Adipose Derived Mesenchymal Stromal Cells Partly through RhoA/ROCK1, but Not ROCK2 Signaling Pathway

doi: 10.1371/journal.pone.0146646

Figure Lengend Snippet: (A) QRT-PCR (up) and western blot (down) analysis of RhoA expression in ADMSCs on day 0, 5 and 10 during the transdifferentiation induction. (B) Predicted binding site between 3’UTR of RhoA mRNA and miR-124 and the designed mutant sequence. (C) Dual luciferase assay of relative luciferase activity of pGL3-RhoA-WT (RhoA-WT) and pGL3-RhoA-MUT (RhoA-WT) transfected with 50 nM miR-124 mimics or the negative control in HEK 293T cells. Firefly luciferase activity was normalized to that of Renilla luciferase. (D) QRT-PCR (up) and western blot (down) analysis of RhoA expression in ADMSCs infected with the pLV-miR-124 locker lentiviral particles or the negative control. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: The ready-to-use human RhoA shRNA lentiviral particles (sc-44209-V), ROCK1 shRNA lentiviral particles (sc-29473-V) and ROCK2 shRNA lentiviral particles (sc-29474-V) were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Binding Assay, Mutagenesis, Sequencing, Luciferase, Activity Assay, Transfection, Negative Control, Infection

Journal: PLoS ONE

Article Title: MiR-124 Promote Neurogenic Transdifferentiation of Adipose Derived Mesenchymal Stromal Cells Partly through RhoA/ROCK1, but Not ROCK2 Signaling Pathway

doi: 10.1371/journal.pone.0146646

Figure Lengend Snippet: (A) QRT-PCR analysis of RhoA, ROCK1 and ROCK2 mRNA expression in ADMSCs infected with the RhoA shRNA lentiviral particles, ROCK1 shRNA lentiviral particles, ROCK2 shRNA lentiviral particles or the negative control. (B) QRT-PCR analysis of NSE, Tuj-1 and GFAP mRNA expression in ADMSCs infected with the pLV-miR-124 locker lentiviral particles, co-infected with the pLV-miR-124 locker lentiviral particles and the RhoA shRNA lentiviral particles, ROCK1 shRNA lentiviral particles or ROCK2 shRNA lentiviral particles on day 1 after transdifferentiation induction. (C, E and G) Representative images of flow cytometry analysis of neurogenic markers (NSE, Tuj-1 and GFAP) positive ADMSCs infected with the pLV-miR-124 locker lentiviral particles and co-infected with the pLV-miR-124 locker lentiviral particles and the RhoA shRNA lentiviral particles (C), ROCK1 shRNA lentiviral particles (E) or ROCK2 shRNA lentiviral particles (G) on day 1 after transdifferentiation induction. (D, F and H) Quantification of the proportion of ADMSCs with positive neurogenic markers showed in C, E and G. *: comparison with shRNA NC or vector NC; #: comparison with transdiff. ADMSCs/miR-124(-). * and # p <0.05, ** and ## p <0.01, *** and ### p <0.001.

Article Snippet: The ready-to-use human RhoA shRNA lentiviral particles (sc-44209-V), ROCK1 shRNA lentiviral particles (sc-29473-V) and ROCK2 shRNA lentiviral particles (sc-29474-V) were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Expressing, Infection, shRNA, Negative Control, Flow Cytometry, Comparison, Plasmid Preparation

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Rho Kinase Type 1 (ROCK1) Promotes Lipopolysaccharide-induced Inflammation in Corneal Epithelial Cells by Activating Toll-Like Receptor 4 (TLR4)-Mediated Signaling

doi: 10.12659/MSM.907277

Figure Lengend Snippet: Influence of ROCK1 on the expression and distribution of RhoA in LPS-treated HCECs. ( A ) Expression and distribution of ROCK1. ( B ) Expression and distribution of RhoA. Scale bar, 100 μm. Each assay was represented by three replicates.

Article Snippet: Antibodies against RhoA (cat. no. A00207), ROCK1 (cat. no. BM4203), and TLR4 (cat. no. BA1717) were purchased from Boster (China).

Techniques: Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Rho Kinase Type 1 (ROCK1) Promotes Lipopolysaccharide-induced Inflammation in Corneal Epithelial Cells by Activating Toll-Like Receptor 4 (TLR4)-Mediated Signaling

doi: 10.12659/MSM.907277

Figure Lengend Snippet: Influence of ROCK1 on the activity of TLR4-mediated NF-κB and ERK signaling in LPS-treated HCECs. ( A ) ROCK1, RhoA, and members of TLR4-mediated NF-κB signaling. ( B ) Members of TLR4-mediated ERK signaling. Each western blotting assay was represented by three replicate and data were expressed as mean ± standard deviation.

Article Snippet: Antibodies against RhoA (cat. no. A00207), ROCK1 (cat. no. BM4203), and TLR4 (cat. no. BA1717) were purchased from Boster (China).

Techniques: Activity Assay, Western Blot, Standard Deviation

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Rho Kinase Type 1 (ROCK1) Promotes Lipopolysaccharide-induced Inflammation in Corneal Epithelial Cells by Activating Toll-Like Receptor 4 (TLR4)-Mediated Signaling

doi: 10.12659/MSM.907277

Figure Lengend Snippet: Relative expression levels of proteins on western blotting assays.

Article Snippet: Antibodies against RhoA (cat. no. A00207), ROCK1 (cat. no. BM4203), and TLR4 (cat. no. BA1717) were purchased from Boster (China).

Techniques: Expressing, Western Blot, Control, Clinical Proteomics