ARHGAP11A Antibody - #DF4425

Figure 1 The levels of ArhGAP11A are significantly increased in patients with AD and APP/PS1 mice A) The relative mRNA levels of ArhGAP11A in the hippocampus of patients with AD and healthy aging people (control [Con]) were analyzed. Data are mean ± SEM, and Mann-Whitney test was used for statistical analysis. (B) The levels of ArhGAP11A in the plasma of patients with MCI and AD. The concentrations of ArhGAP11A in the plasma of patients clinically diagnosed as MCI, patients with AD, and healthy people were detected by ELISA kit. n = 16–17 samples per group. Data are mean ± SEM, and Kruskal-Wallis one-way ANOVA with two-stage step-up method Benjamini, Krieger, and Yekutieli test was used for statistical analysis. (C) The levels of ArhGAP11A in the brain homogenates of 4- to 12-month-old APP/PS1 mice and wild-type (WT) mice were detected by western blotting. (D) Densitometry analysis of ArhGAP11A in (C). n = 4 mice per group. Data are mean ± SEM, and a two-way ANOVA followed by Tukey’s multiple comparison test was used for statistical analysis. (E) Representative confocal images of ArhGAP11A (green) in the cortical neurons (red) of 4- to 12-month-old APP/PS1 mice and WT mice. Scale bar represents 5 μm. (F) The fluorescent area of ArhGAP11A in (E) was quantified by ImageJ software. n = 5 mice per group. Data are mean ± SEM, and a two-way ANOVA followed by Tukey’s multiple comparison test was used for statistical analysis. For (C) and (E), data are representative of three independent experiments with similar results, and for (D) and (F), data are pooled from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns, not significant. See also Figures S1 and S7 and Table S1.

Figure 1 The levels of ArhGAP11A are significantly increased in patients with AD and APP/PS1 mice A) The relative mRNA levels of ArhGAP11A in the hippocampus of patients with AD and healthy aging people (control [Con]) were analyzed. Data are mean ± SEM, and Mann-Whitney test was used for statistical analysis. (B) The levels of ArhGAP11A in the plasma of patients with MCI and AD. The concentrations of ArhGAP11A in the plasma of patients clinically diagnosed as MCI, patients with AD, and healthy people were detected by ELISA kit. n = 16–17 samples per group. Data are mean ± SEM, and Kruskal-Wallis one-way ANOVA with two-stage step-up method Benjamini, Krieger, and Yekutieli test was used for statistical analysis. (C) The levels of ArhGAP11A in the brain homogenates of 4- to 12-month-old APP/PS1 mice and wild-type (WT) mice were detected by western blotting. (D) Densitometry analysis of ArhGAP11A in (C). n = 4 mice per group. Data are mean ± SEM, and a two-way ANOVA followed by Tukey’s multiple comparison test was used for statistical analysis. (E) Representative confocal images of ArhGAP11A (green) in the cortical neurons (red) of 4- to 12-month-old APP/PS1 mice and WT mice. Scale bar represents 5 μm. (F) The fluorescent area of ArhGAP11A in (E) was quantified by ImageJ software. n = 5 mice per group. Data are mean ± SEM, and a two-way ANOVA followed by Tukey’s multiple comparison test was used for statistical analysis. For (C) and (E), data are representative of three independent experiments with similar results, and for (D) and (F), data are pooled from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns, not significant. See also Figures S1 and S7 and Table S1.

Figure 2. ARHGAP11A is highly expressed in lung adenocarcinoma. (A) ARHGAP11A showed prominently high expression in tumor tissue than in non-cancerous tissues based on Wilcoxon rank-sum test from GEO database. (B) The protein expression levels of ARHGAP11A based on CPTAC. (C) Validation of protein expression levels of ARHGAP11A in the HPA database. Tumor tissue, https://www.proteinatlas.org/ENSG00000198826-ARHGAP11A/pathology/lung+cancer#img; normal tissue, https://www.proteinatlas.org/ENSG00000070193-FGF10/tissue/lung#img. (D) Expression of ARHGAP11A in human bronchial epithelial cells (BEAS-2B) and lung adenocarcinoma cells (H1299 and A549) by Western blot and using Image J quantitative analysis software. **, P

Figure 4 ARHGAP11A interacts with TPM1 in gastric cancer cells. (a) Western blot analysis verified the overexpression efficiency of ARHGAP11A in gastric cancer cells. (b) Western blot detection after IP. (c) Matching peptides in ARHGAP11A and TPM1. (d) Left: Flag-tagged ARHGAP11A and HA-tagged TPM1 plasmids were co-transfected into HEK293T cells for 36 h followed by cell lysate preparation and IP assay with anti-Flag beads followed by immunoblotting with indicated antibodies. Right: HA-tagged ARHGAP11A and Flag-tagged TPM1 plasmids were cotransfected into HEK293T cells for 36 h followed by cell lysate preparation and IP assay with anti-Flag beads followed by immunoblotting with indicated antibodies. IP: immunoprecipitates, WCL: whole-cell lysates. (e) The interaction of ARHGAP11A and TPM1 was tested in AGS cells. AGS cells overexpressing Flag-ARHGAP11A were lysed with cell lysate, followed by IP assay with anti-Flag beads followed by immunoblotting with indicated antibodies. (f) ARHGAP11A domain. (g) Analysis of the domain involved in the interaction between ARHGAP11A and TPM1. HEK293T cells were transiently cotransfected with plasmids expressing Flag-tagged of indicated ARHGAP11A mutant plasmids and HA-tagged TPM1 plasmids, followed by cell lysate preparation and IP assay with anti-Flag beads followed by immunoblotting with indicated antibodies. IP: immunoprecipitates, WCL: whole-cell lysates.