anti arp3 a 1  (Santa Cruz Biotechnology)

Journal: Nature Communications

Article Title: Compartmentalized cytoplasmic tradewinds direct soluble proteins

doi: 10.1038/s41467-026-70688-6

Figure Lengend Snippet: a Asymmetry analysis of spot activation of mEos3.2 Arp3, vinculin, and paxillin in CAD cells illustrates that they are targeted toward the leading edge, where polymerization and adhesions initiate ( n = 14, Arp3, 9 vinculin, 10 paxillin cells). b Temporal color-coded spot activation of mEos3.2 actin in an NG108 cell advancing preferentially on the right illustrates that this is the region where recently activated actin is targeted (arrows). c Maximum projection of 3D SIM images of Alexa 647-phalloidin and mEmerald MLC in control cells and cells treated with 0.2 μM nitro-blebbistatin before fixation illustrates the broadening and flattening of the actin-myosin arcs at the border between the lamella and cell body when myosin is inhibited. d These observations were quantified by fitting the arcs to ellipses and calculating ellipticity, which reveals flatter arcs in the blebbistatin-treated cells ( n = 19 control cells, 87 measurements, 17 blebbistatin cells, 100 measurements). Ellipticity =√(1-(minor) 2 major −2 )) e An overlay of two different mNeon MLC image time points ( t = 0, and t = 45 s) with arrows indicating where the edge retracts and extends during the 45 s. f Contour of the same two-time points in ( e ) with arrows indicating changes in the position and curvature of the arcs corresponding to the retraction and advancement of the leading edge. g CAD cell expressing mEmerald MLC were treated with a high-power localized beam of 405 light positioned over one arc, causing the arc to be disrupted and only the leading edge in front of that single arc to collapse. The yellow line indicates the position of the edge prior to MLC disruption. p values two-tailed t -tests. Box plots show median (center line), interquartile range (25th–75th percentiles; box), whiskers to the most extreme non-outlier values, and notches as 95% CIs. Scale bars, 5 μm. Source data are provided as a Source Data file.

Article Snippet: Antibodies used were: anti-actin (Sigma, A5441), anti-paxillin (Sigma, P1093), GFP (Abcam, AB38689), GAPDH (Applied Bioscience, AM4300), Eos (Novus, NP3-05557), Arp3 (Proteintech, 13822), and Vinculin (Sigma, V9131).

Techniques: Activation Assay, Control, Expressing, Disruption, Two Tailed Test

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: The expression of ACTR3 is notably elevated in various cancers, including CESC. (A) The relationship between the expression levels of ACTR3 and OS in patients with various cancer types was assessed through univariate Cox regression analysis. (B) The radar chart depicts the expression levels of ACTR3 in tumor tissues. (C) A heatmap analysis was conducted to evaluate the correlation between the expression levels of ACTR3 and OS in patients with various cancer types. (D) The expression levels of ACTR3 in various human cancer tissues were analyzed in comparison to those in normal tissues, utilizing data sourced from the TCGA and GTEx databases. GTEx = genotype-tissue expression, OS = overall survival, TCGA = The Cancer Genome Atlas.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Expressing, Comparison

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: Expression of ACTR3-related genes and their prognostic significance. (A) A schematic representation illustrating the relationships among ACTR3 and several proteins obtained from the STRING database. (B) The forest plot illustrates the significance of genes associated with ACTR3 in terms of their predictive value. (C) The comparison of mRNA expression levels of ACTR3-related genes was conducted between normal tissues and tumor tissues.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Expressing, Comparison

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: Analysis of functional enrichment concerning DEGs predicated on the expression levels of ACTR3. (A–C) The GO enrichment analysis conducted on DEGs associated with ACTR3 identified significant enrichment in various biological processes, cellular components, and molecular functions. (D) An analysis of KEGG pathway enrichment pertaining to ACTR3 in CESC was conducted. (E–F) The grid chart presents the results obtained from the GO and KEGG analyses. CESC = cervical squamous cell carcinoma and endocervical adenocarcinoma, DEGs = differential expressed genes, GO = gene ontology, KEGG = Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Functional Assay, Expressing

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: GSEA enrichment analysis of ACTR3. (A) The heat maps illustrate the ten genes that exhibit a positive co-expression relationship with ACTR3. (B) The heat maps illustrate the ten genes that exhibit a negative co-expression relationship with ACTR3. (C) The results obtained from GSEA are illustrated using mountain plots. (D–G) An examination of the functional roles and pathway enrichment related to ACTR3 is provided. GSEA = gene set enrichment analysis.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Expressing, Functional Assay

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: The examination of the relationship between the infiltration of immune cells and the expression levels of ACTR3. (A) Spearman correlation analysis was conducted to evaluate the relationship between the infiltration levels of 24 distinct immune cell types and the expression levels of ACTR3. (B–L) A scatter plot illustrating the extent of immune cell infiltration in relation to varying levels of ACTR3 expression. (M) A heatmap depicting the infiltration of immune cells corresponding to different levels of ACTR3 expression.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Expressing

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: The influence of ACTR3 expression on prognosis and its diagnostic significance. (A) The relationship between OS and the expression levels of ACTR3 across different clinical subgroups of CESC. (B) Proportion of mortality as risk score values escalated within low and high-risk groups. (C) Time-dependent survival ROC curves were generated to estimate the survival probabilities at 1-, 3-, and 5-yr intervals for patients diagnosed with CESC, utilizing the expression levels of ACTR3 as the predictive variable. (D) A nomogram was constructed to forecast the OS rates at 1, 3, and 5 yr for individuals diagnosed with CESC. CESC = cervical squamous cell carcinoma and endocervical adenocarcinoma, OS = overall survival, ROC = receiver operating characteristic.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Expressing, Diagnostic Assay, Generated, Construct

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: Association between ACTR3 expression and clinical features. (A–G) The correlation between ACTR3 expression and multiple clinical parameters – such as the T stage, N stage, M stage, OS events, DSS events, PFI events, and histological type – have been explored. (H–J) Calibration plots were generated to assess the accuracy of predictions for OS at 1-, 3-, and 5-yr intervals. (K) A nomogram has been developed to forecast the 1-, 3-, and 5-yr OS rates in patients diagnosed with CESC. CESC = cervical squamous cell carcinoma and endocervical adenocarcinoma, DSS = disease-specific survival, OS = overall survival, PFI = progression-free interval.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Expressing, Generated

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: Analysis of single-cell RNA sequencing data from CESC tissues. (A and B) The bar chart, together with the stacked bar chart, depicts the cellular quantities observed in both the control group and the tumor group. (C and D) The allocation of various cell types among the control and tumor groups. (E) The PCA, t-SNE, and UMAP techniques were employed to visualize the 18 unique cellular clusters. (F, G) The clusters received further annotation through the use of designated marker genes. (H) The UMAP visualization illustrates the expression levels of ACTR3 distributed among various cell clusters. (I) Violin plots illustrating the expression levels of ACTR3 across all recognized cell types. (J) The expression of ACTR3 across all recognized cell types. CESC = cervical squamous cell carcinoma and endocervical adenocarcinoma.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: RNA Sequencing, Control, Marker, Expressing

Journal: Medicine

Article Title: Expression of ACTR3 in cervical cancer and impact on immune cell infiltration and prognosis: A comprehensive analysis based on bulk RNA-Seq and single-cell RNA-Seq

doi: 10.1097/MD.0000000000046316

Figure Lengend Snippet: Validation of ACTR3 expression in clinical specimens from patients diagnosed with CESC. (A) The protein expression levels of ACTR3, KI67, and CK7 were found to be significantly higher in CESC compared to those in normal tissue samples. (B) Statistical analysis of ACTR3, KI67, and CK7 positive cells. CESC = cervical squamous cell carcinoma and endocervical adenocarcinoma.

Article Snippet: Subsequently, the sections were exposed to primary antibodies, specifically ACTR3 (dilution of 1:1000; Proteintech, Wuhan, China) and KI67, CK7, p40, p16, and p63 (dilution of 1:1000; Zhongshan Golden Bridge Biotechnology Co., Beijing, China), for an overnight incubation at 4°C.

Techniques: Biomarker Discovery, Expressing

Journal: bioRxiv

Article Title: Brush border intermicrovillar adhesion limits bacteria attachment to the small intestine brush border

doi: 10.1101/2025.11.25.690363

Figure Lengend Snippet: (A) Single confocal plane at the villus surface of Cdhr2 Δepi mouse ileum whole mount tissue stained with DAPI (DNA). Boxed areas enlarged (zooms 1-3) with additional stains DAPI (DNA), phalloidin (F-actin), EPS8, and F-actin/EPS8 overlay. Adherent bacteria marked with a transparent overlay (yellow, single bacteria; blue, filaments). White arrowheads highlight EPS8 localization in enterocytes (zoom 1). Teardrop-shaped bacteria indicated by black arrowheads (zooms 1-2). (B) Cdhr2 Δepi mouse ileum frozen tissue section stained with DAPI (DNA), deconvolved, AI segmented to isolate and differentially contrast bacterial DNA and viewed as a maxIP (depth 12.6 µm). The bacteria channel is z-depth color-coded; closer bacteria purple/magenta, deeper bacteria yellow/white, z-depth color-code at right. DNA from cell nuclei (cyan/blue). Boxed areas enlarged below (zooms 1-3) with the box color indicating the z-depth of the zoom images. (B, zoom 1) IRTKS localization in the enterocyte brush border. (B, zooms 2-3) highlights adherent bacteria. Column 1 is the main image panel enlarged (maxIP, depth 12.6 µm). Subsequent columns 2-4 show additional stains phalloidin (F-actin), IRTKS, and F-actin/IRTKS overlay as a maxIP (depth 0.3 µm). White boxed areas within zoom images (columns 2-4) are contrast enhanced and shown as insets (upper left) with bacteria localization indicated by ovals (yellow, single bacteria; blue, filaments). (C-E) Single confocal planes at the villus surface of Cdhr2 Δepi mouse ileum whole mount tissue stained with DAPI (DNA). Boxed areas (zooms 1-2) enlarged at right. DAPI (DNA) and phalloidin (F-actin) are shown in all sets plus Villin (VIL1, C), F-actin/VIL1 (C), Espin (ESPN, D), and Arp3 (ACRT3, E). (C, zoom 1), black arrowheads indicate a filamentous attachment with eccentric VIL1 localization. (F) Cdhr2 Δepi mouse ileum paraffin tissue section stained for DAPI (DNA) and actin, deconvolved, and viewed as a maxIP (depth 10.4 µm). A z-depth color-code is applied to each channel, scales lower right. DNA AI segmented to isolate bacteria, cell nuclei are not displayed. Boxed areas enlarged at right (zooms 1-3). Column 1, bacteria/actin maxIP (depth 10.4 µm) enlarged from the main panel. Column 2, intensity color-coded SFB FISH (SFB) maxIP (depth 10.4 µm), intensity scale at right). Boxed areas in zoom 1 further enlarged in zoom 4. Z-depth indicated by box color, based on the main panel bacteria z-depth color-code. (F, zoom 4) Single confocal plane of surface bacteria with additional stains; DAPI (DNA) without AI segmentation, SFB FISH (SFB) intensity color-coded, actin/EPS8 overlay, and actin/EPS8 plus DNA overlay. White arrowheads indicate SFB FISH negative bacteria. (G) SEM images of Cdhr2 Δepi mouse terminal ileum. Boxed areas enlarged in zooms 1-4 to highlight adherent bacteria (yellow, single bacteria; blue, filaments). (G, zooms 1, 3) single bacteria. (G, zooms 2, 4), filamentous bacteria with elongated microvilli at the base. Scale bars: 20 µm (A-F, main panels), (B, F) z-depth color-code tick marks at 1 µm intervals, 5 µm (A-F zooms 1-3), 2.9 µm (B, zoom 1-3, insets), 1 µm (F, zoom 4), 10 µm (G, main panel), 2 µm (G, zooms 1-4).

Article Snippet: The following antibodies and dilutions were used for whole mount staining: anti-EPS8 (mouse BD Transduction Laboratories #610144), 1:400; anti-Villin (rabbit, Santa Cruz #sc-28283), 1:50; anti-ESPN (rabbit, Sigma-Aldrich #HPA028674), 1:500; anti-Arp3 (ACTR3) (mouse, Santa Cruz #sc-48344), 1:100; Goat anti-mouse Alexa Fluor 488 F(ab’)2 fragment (Invitrogen #A-11017), 1:1000; Goat anti-rabbit Alexa Fluor 568 F(ab’)2 fragment (Invitrogen #A-21069), 1:1000; Phalloidin 647 (Invitrogen #A22287), 1:200; and DAPI (ThermoFisher #62248), 1:200; and Hoechst 33342 (ThermoFisher #62249), 1:200.

Techniques: Staining, Bacteria

Journal: Cell Communication and Signaling : CCS

Article Title: Semaphorin 5A modulates focal adhesion pathway and lamellipodia formation in melanoma

doi: 10.1186/s12964-025-02526-z

Figure Lengend Snippet: Regulation of lamellopodia by SEMA5A. A Western blot and relative densitometric analyses of Arp3 protein in total protein extracts of A375 control (shCTRL) and SEMA5A knockdown (sh5A#2 and #4) clones, plated on Collagen Type I (4 h). One representative western blot analysis out of three with similar results is reported. GAPDH was used as loading and transfer control. The data represent the ratio (mean ± SEM) of the protein band intensity normalized to GAPDH in the silenced conditions versus the control from three independent biological repeats. B Confocal laser scanning microscopy analysis of A375 shCTRL or sh5A#2 cells, plated on Collagen Type I for 4 h. Cells were stained for Arp3 (green), F-actin by phalloidin staining (red), nuclei are in blue (DAPI). Arrows indicate Arp3 positive lamellipodia. The zoomed area indicates the region of actin/Arp3 nucleation. C Bar chart showing Person’s correlation coefficient (± SEM) calculated for Arp3 (green) and F-actin (red) in A375 shCTRL and sh5A#2 using the Coloc2 Plugin of ImageJ. A,C Statistical analysis was performed applying unpaired two-tailed student’s t test, *p < 0.05Regulation of lamellopodia by SEMA5A. A Western blot and relative densitometric analyses of Arp3 protein in total protein extracts of A375 control (shCTRL) and SEMA5A knockdown (sh5A#2 and #4) clones, plated on Collagen Type I (4 h). One representative western blot analysis out of three with similar results is reported. GAPDH was used as loading and transfer control. The data represent the ratio (mean ± SEM) of the protein band intensity normalized to GAPDH in the silenced conditions versus the control from three independent biological repeats. B Confocal laser scanning microscopy analysis of A375 shCTRL or sh5A#2 cells, plated on Collagen Type I for 4 h. Cells were stained for Arp3 (green), F-actin by phalloidin staining (red), nuclei are in blue (DAPI). Arrows indicate Arp3 positive lamellipodia. The zoomed area indicates the region of actin/Arp3 nucleation. C Bar chart showing Person’s correlation coefficient (± SEM) calculated for Arp3 (green) and F-actin (red) in A375 shCTRL and sh5A#2 using the Coloc2 Plugin of ImageJ. A , C Statistical analysis was performed applying unpaired two-tailed student’s t test, *p < 0.05

Article Snippet: Primary antibodies were: Arp3 (#A4514), FAK (#A11131) from Abclonal (Düsseldorf, Germany), phosphorylated FAK (Y567, # 2H74L24) from Invitrogen (ThermoFisher), active Integrin β1 chain (#MAB2079Z) from Merck.

Techniques: Western Blot, Control, Knockdown, Clone Assay, Confocal Laser Scanning Microscopy, Staining, Two Tailed Test