ha arhgap29  (Addgene inc)

Journal: Oncology reports

Article Title: ARHGAP29 expression may be a novel prognostic factor of cell proliferation and invasion in prostate cancer.

doi: 10.3892/or.2020.7811

Figure Lengend Snippet: Figure 1. AR, YAP, ARHGAP29, and F‑actin expression in prostate cancer cell lines. (A) mRNA levels of ARHGAP29 in prostate cancer cell lines (22Rv1, LNCaP, DU145 and PC‑3). Experiments were performed in triplicate. The vertical axis of the graph is presented on a logarithmic scale. The results are expressed as the mean ± SD. *P<0.01 compared with PC‑3 cells. (B) Western blotting of the expression of various proteins in prostate cancer cell lines. (C) Densitometric analysis of B (relative protein expression to GAPDH). There was an inverse association between ARHGAP29 and F‑actin expression, but no clear association between the expression of ARHGAP29 and that of other proteins. The results are expressed as the mean ± SD (at least three independent experiments). *P<0.05, **P<0.01 compared with PC‑3 cells. AR, androgen receptor; YAP, yes‑associated protein; ARHGAP29, Rho GTPase‑activating protein 29.

Article Snippet: A mammalian expression of HA tagged ARHGAP29 (#104154) was purchased from Addgene, Inc.

Techniques: Expressing, Western Blot

Journal: Oncology reports

Article Title: ARHGAP29 expression may be a novel prognostic factor of cell proliferation and invasion in prostate cancer.

doi: 10.3892/or.2020.7811

Figure Lengend Snippet: Figure 2. Effects of downregulation or upregulation of ARHGAP29 in prostate cancer cell lines (PC‑3, LNCaP and DU145) on the mRNA and protein expres- sion. (A) Quantitative comparison of ARHGAP29 expression by RT‑qPCR in prostate cancer cell lines with downregulation or upregulation of ARHGAP29 compared with the control. Experiments were performed in triplicate. The results are expressed as the mean ± SD. *P<0.01 compared with si‑NC (PC‑3 cells) or pcDNA empty vector (mock; LNCaP and DU145 cells). In LNCaP and DU145, the vertical axis of the graph is presented on a logarithmic scale. (B) Western blotting of the expression of various proteins in prostate cancer cell lines. (C) Densitometric analysis of B (relative protein expression to GAPDH). The protein expression was only slightly altered in ARHGAP29‑upregulated LNCaP and DU145 cells compared with ARHGAP29‑downregulated PC‑3 cells. The results are expressed as the mean ± SD (at least three independent experiments). *P<0.01 compared with si‑NC (PC‑3 cells) or pcDNA empty vector (mock; LNCaP and DU145 cells). (D) The ratio of phosphorylated protein to non‑phosphorylated protein (YAP and cofilin). The results are expressed as the mean ± SD (at least three independent experiments). *P<0.05, **P<0.01 compared with si‑NC (PC‑3 cells) or pcDNA empty vector (mock; LNCaP and DU145 cells). ARHGAP29, Rho GTPase‑activating protein 29.

Article Snippet: A mammalian expression of HA tagged ARHGAP29 (#104154) was purchased from Addgene, Inc.

Techniques: Comparison, Expressing, Control, Plasmid Preparation, Western Blot

Journal: Oncology reports

Article Title: ARHGAP29 expression may be a novel prognostic factor of cell proliferation and invasion in prostate cancer.

doi: 10.3892/or.2020.7811

Figure Lengend Snippet: Figure 3. Effects of downregulation or upregulation of ARHGAP29 in prostate cancer cell lines (PC‑3, LNCaP and DU145) on cell viability and invasion. (A) MTS assay results. Cell viability was significantly decreased in ARHGAP29‑downregulated PC‑3 cells compared with control cells (si‑NC). In contrast, cell viability was significantly increased in ARHGAP29‑upregulated LNCaP and DU145 cells compared with control cells (mock). Experiments were per- formed in triplicate. The results are expressed as the mean ± SD. *P<0.05, **P<0.01 compared with si‑NC (PC‑3 cells) or pcDNA empty vector (mock; LNCaP and DU145 cells). (B) Invasion assay results. The number of invasive PC‑3 cells was significantly decreased after knocking down ARHGAP29. In contrast, the number of invasive LNCaP and DU145 cells was significantly increased after upregulation of ARHGAP29. Experiments were performed in triplicate. The results are expressed as the mean ± SD. *P<0.01 compared with si‑NC (PC‑3 cells) or pcDNA empty vector (mock; LNCaP and DU145 cells). ARHGAP29, Rho GTPase‑activating protein 29.

Article Snippet: A mammalian expression of HA tagged ARHGAP29 (#104154) was purchased from Addgene, Inc.

Techniques: MTS Assay, Control, Plasmid Preparation, Invasion Assay

Journal: Oncology reports

Article Title: ARHGAP29 expression may be a novel prognostic factor of cell proliferation and invasion in prostate cancer.

doi: 10.3892/or.2020.7811

Figure Lengend Snippet: Figure 4. Identification of other cell motility‑related genes after knockdown of ARHGAP29. (A) The 84 genes related to cell motility are presented and analysis of the results was performed using the web‑based software ‘RT2 Profiler PCR Array’ Data Analysis version 3.5. All of genes involved in the array are presented. (B) A heatmap is presented. Numerous genes were downregulated after knockdown of ARHGAP29 in PC3 cells. A1‑G12 corresponds to A1‑G12 of A. In the heatmap the data was displayed in a grid where each row represents a gene included in the commercial array. The color and intensity of the boxes represent changes of gene expression. For example, red represents upregulated genes and blue represents downregulated genes. (C) PCR Array of cell motility genes in PC‑3 cells before and after downregulation of ARHGAP29. The graph shows the relative log value between cell motility gene expression in PC‑3 cells before and after downregulation of ARHGAP29 (vertical axis, after downregulation of ARHGAP29; horizontal axis, before downregulation of ARHGAP29). The diagonal line in the center of the graph shows equal level of gene expression in PC‑3 cells before and after downregulation of ARHGAP29. The upper diagonal lines indicate that the level of gene expression after downregulation was three times higher than that of before downregulation. The lower diagonal line indicates that the level of gene expression after downregulation was one third times lower than that of before downregulation. The mRNA level of MMP‑2 was significantly downregulated. (D) Quantitative comparison of MMP‑2 by RT‑qPCR between si‑NC and si‑ARHGAP29 PC‑3 transfectants. MMP‑2 mRNA expression was suppressed after downregulation of ARHGAP29. Experiments were performed in triplicate. The results represent the mean ± SD. *P<0.01 compared with si‑NC. ARHGAP29, Rho GTPase‑activating protein 29.

Article Snippet: A mammalian expression of HA tagged ARHGAP29 (#104154) was purchased from Addgene, Inc.

Techniques: Knockdown, Software, Gene Expression, Comparison, Expressing

Journal: Oncology reports

Article Title: ARHGAP29 expression may be a novel prognostic factor of cell proliferation and invasion in prostate cancer.

doi: 10.3892/or.2020.7811

Figure Lengend Snippet: Figure 5. Association between the expression levels of YAP and ARHGAP29 and the prognosis of prostate cancer patients. (A) Histology (H&E staining) and IHC staining of YAP or ARHGAP29. Protein expression patterns were different between YAP and ARHGAP29. YAP was heterogeneously stained. Scale bars represent 200 µM. (B) Association of the D’Amico risk classification with the expression of YAP and ARHGAP29. When comparing three groups, one‑way ANOVA (inside the black frame) followed by Tukey‑Kramer test were used. *P<0.05. YAP was unrelated to the risk classification, but ARHGAP29 was signifi- cantly associated with the risk classification. The diamond indicates the mean (long horizontal line) and 95% confidence interval of the H‑score. (C) ROC curve of YAP, ARHGAP29 and both (AUC: 0.5971, 0.6216 and 0.6400, respectively). Both proteins had low AUC scores as prognostic markers. (D) Kaplan‑Meier plot of biochemical PFS stratified by the expression levels of YAP, ARHGAP29 and both. For each protein, high expression was associated with a poor prognosis of prostate cancer patients. Furthermore, the group with high expression of both YAP and ARHGAP29 had the worst prognosis. PFS was compared by a log‑rank test. P<0.05 was considered to indicate a statistically significant difference. YAP, yes‑associated protein; ARHGAP29, Rho GTPase‑activating protein 29; ROC, receiver operating characteristic; AUC, area under the curve; PFS, progression‑free survival; NS, not significant.

Article Snippet: A mammalian expression of HA tagged ARHGAP29 (#104154) was purchased from Addgene, Inc.

Techniques: Expressing, Staining, Immunohistochemistry