Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 1 Abnormally high expression of ARPC5 in glioma. (A) Differential expression of ARPC5 in various tumor types was analyzed based on the TIMER online website. (B) The distinct upregulation of ARPC5 in glioma was demonstrated in GEPIA online website. (C) Box plot based on the expression of ARPC5 in the GSE2223 (Glioma = 50, Normal = 4). (D) Box plot based on the expression of ARPC5 in the GSE29796 (Glioma = 52, Normal = 20). (E) Box plot based on the expression of ARPC5 in the GSE116520 (Glioma = 34, Normal = 8). (F) Box plot based on the protein expression of ARPC5 in the CPTAC samples (Glioma = 99, Normal = 10). (G) ARPC5 protein expression was detected in cerebral cortex (Patient id: 1582), low grade glioma (Patient id: 3365), and high-grade glioma (Patient id: 3251) tissues from HPA dataset. (H) Representative images of ARPC5 expression in glioma and its surrounding tissues. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Expressing, Quantitative Proteomics

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 2 Correlation between different status of ARPC5 expression and prognosis of glioma patients. (A) Survival analysis of ARPC5 in CGGA database. (B) ROC curve analysis of ARPC5 at 1, 3, and 5 years in CGGA database. (C) Survival analysis of ARPC5 in TCGA database. (D) ROC curve analysis of ARPC5 at 1, 3, and 5 years in TCGA database. Survival analysis of the signature in patients stratified by grade (E, F), gender (G, H), age (I, J), IDH mutation status (K, L), 1p/19q codeletion status (M, N), and MGMTp methylation status (O, P) in CGGA database

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Expressing, Mutagenesis, Methylation

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 3 Prognostic significance of ARPC5 and its association with drug sensitivity in glioma. (A) Univariate regression analysis of prognosis in CGGA data base. (B) Multivariate analysis of prognosis in CGGA database. (C) The nomogram to predict the association between ARPC5 expression and OS was de veloped using the CGGA dataset. (D) The calibration curve for the nomogram-predicted OS. ARPC5 affects the sensitivity to 5-fluorouracil (E), bleomycin (F), etoposide (G), crizotinib (H), sorafenib (I), and erlotinib (J)

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Expressing

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 4 Enrichment analysis identifies ARPC5-associated signaling pathways. (A) Heatmap for DEGs generated by compare the difference of ARPC5 ex pression in glioma from the CGGA dataset. (B) Volcano plots illustrated all DEGs. Barplot showing the GO (C) and KEGG (D) analysis for ARPC5 in glioma. (E) GO functional annotation of ARPC5 in glioma from GSEA. (F) KEGG pathway analysis of ARPC5 in glioma from GSEA.

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Protein-Protein interactions, Generated, Functional Assay

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 5 Investigations the expression of ARPC5 in glioma through single-cell analysis. (A) Through dimension reduction analysis of CGGA single-cell data, TSNE plot depicted that 6,148 cells were classified into 16 clusters. (B) The violin plot showed the expression of ARPC5 in each type of cluster. (C) Sixteen clusters were divided into 5 types of cells: astrocyte, macrophage, monocyte, epithelial and T cell. (D) The scatter plot shows the expression of ARPC5 in each cell

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Expressing, Single-cell Analysis

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 6 Correlations of ARPC5 expression with immunity-related indexes. (A) Violin plot showed the correlation between immune classification and tumor purity. The immune infiltration models of low- and high-ARPC5 were detected through ssGSEA methods in glioma from the TCGA database. (C) Violin plot showed the correlation between ARPC5 with stromal scores, immune scores, and ESTIMATE scores. (D) Lollipop showed the correlation between ARPC5 with infiltrating immune cells. Scatter plot showed the correlation between ARPC5 with TMB (E) and MSI (F). (G-J) Violin plot showed the correla tion between ARPC5 with immunotherapy

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Expressing

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 7 Effect of ARPC5 on the expression of CD3 and prognosis. (A) Representative images revealed the relationships between the ARPC5 expression and T cell marker CD3 in gliomas. (B) Survival analysis of ARPC5 in glioma chip ZL-BraG180sur01.

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Expressing, Marker

Journal: BMC cancer

Article Title: ARPC5 acts as a potential prognostic biomarker that is associated with cell proliferation, migration and immune infiltrate in gliomas.

doi: 10.1186/s12885-023-11433-w

Figure Lengend Snippet: Fig. 8 Effect of ARPC5 on the proliferation and migration of glioma cells. (A) The expression level of ARPC5 protein was detected in LN229 and U251 cells after shRNA interference, GAPDH was used to confirm equal protein loading. (B) RT-qPCR verified the expression efficiency of ARPC5 in LN229 and U251 cells after shRNA interference. Growth curve assessing the effect of ARPC5 on the proliferation of LN229 (C) and U251 (D) cells. (E) The representa tive image showed the clone formation ability of LN229 and U251 cells after transfection. (F) Transwell analysis was further used to examine the effect of ARPC5 on migration of LN229 and U251 cells, and the number of migrating cells was quantitatively analyzed. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Glioma tissue samples and glioma chip ZLBraG180sur01 were incubated with ARPC5 antibody (16717-1-AP, proteintech, Wuhan, China) and CD3 antibody (17617-1-AP, proteintech, Wuhan, China).

Techniques: Migration, Expressing, shRNA, Quantitative RT-PCR, Transfection

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes

doi: 10.1038/s41374-018-0178-5

Figure Lengend Snippet: [A] Cellular senescence was enhanced in BDL WT mice compared to WT mice but reduced in BDL α-CGRP −/− mouse liver compared to BDL WT mice, which was verified by SA-β-gal staining in liver sections (n=4, Orig., magnification, 40×; scale bar=100μm). [B-C] The mRNA expression of [B] p16, p21 and CCL2 and PAI-1 as well as [C] p16 protein levels were increased in total liver from BDL WT mice compared to WT mice but decreased in total liver from BDL α-CGRP −/− mice compared to BDL WT mice (n=3). * p <0.05 vs. WT mice; # p <0.05 BDL WT mice.

Article Snippet: The mouse antibody against p16 (sc-166760) was purchased from Santa Cruz Biotechnology.

Techniques: Staining, Expressing

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes

doi: 10.1038/s41374-018-0178-5

Figure Lengend Snippet: [A] The mRNA expression of α-SMA, Col1α1 and Fn1 was decreased in cholangiocytes from BDL α-CGRP −/− mice compared to BDL WT mice (n=4). [B] The mRNA expression of p16, p21 and PAI-1 was decreased in cholangiocytes from BDL α-CGRP −/− mice compared to BDL WT mice (n=4). [C] The mRNA expression of SASP markers such as IL-6, IL-8, IL1-β and MMP2 was decreased in cholangiocytes from BDL α-CGRP −/− mice compared to BDL WT mice (n=4). [D] Immunofluorescence demonstrated that p16 protein expression was decreased in cholangiocytes from BDL α-CGRP −/− mice compared to BDL WT mice (n=3, Orig. magnification, 20×, scale bar= 50 μm). * p 0.05 vs. WT mice; # p <0.05 vs. BDL WT mice.

Article Snippet: The mouse antibody against p16 (sc-166760) was purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Immunofluorescence

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes

doi: 10.1038/s41374-018-0178-5

Figure Lengend Snippet: [A] The mRNA expression of Col1α1 was decreased in HSCs isolated from BDL α-CGRP −/− mice compared to BDL WT mice (n=3). [B] The p16 and p21 mRNA expression was increased in BDL α-CGRP −/− mice compared to BDL WT mice (n=3). [C] Immunofluorescent staining showed that p16 protein expression was decreased in HSCs from BDL WT mice compared to BDL α-CGRP −/− mice (n=3, Orig. magnification, 40×, scale bar= 25 μm). # p <0.05 vs. BDL WT mice.

Article Snippet: The mouse antibody against p16 (sc-166760) was purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Isolation, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes

doi: 10.1038/s41374-018-0178-5

Figure Lengend Snippet: [A] Representative immunofluorescence picture of receptor activity-modifying protein 1 (RAMP1) in HHSCs and IMCLs were shown (n=4, Orig., magnification. 40×; scale bar=50μm). [B-C] α-CGRP stimulated the expression of α-SMA and Col1 α 1 in both [B] IMCLs and [C] HHSCs, which was prevented by CGRP 8–37 (n=4). [D] The expression of p16 and p18 was decreased in HHSCs while increased in ICMLs simulated by α-CGRP; these effects were partly reversed by incubation with CGRP 8–37 (n=4). * p <0.05 vs. Basal; # p <0.05 vs. α-CGRP-treated group.

Article Snippet: The mouse antibody against p16 (sc-166760) was purchased from Santa Cruz Biotechnology.

Techniques: Immunofluorescence, Activity Assay, Expressing, Incubation

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes

doi: 10.1038/s41374-018-0178-5

Figure Lengend Snippet: [A-C] The mRNA expression of [A] TGF-β1, [B] TIMP1 and [C ] Fn1 increased in HHSCs stimulated with cholangiocyte supernatant from BDL WT mice compared to HHSCs stimulated with cholangiocyte supernatant from WT mice but decreased in HHSCs treated with cholangiocyte from BDL α-CGRP −/− mice compared to HHSCs stimulated with cholangiocyte supernatant from BDL WT mice (n=4). [D-F] The mRNA expression of [D] p16, [E] p18 and [F] p21 was increased in HHSCs treated with cholangiocyte supernatant from BDL α-CGRP −/− mice compared to HHSCs stimulated with BDL cholangiocyte supernatant (n=4). * p <0.05 vs. HHSCs treated with cholangiocyte supernatant from WT mice; # p <0.05 vs. HHSCs treated with cholangiocyte supernatant from BDL WT mice.

Article Snippet: The mouse antibody against p16 (sc-166760) was purchased from Santa Cruz Biotechnology.

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Feedback-Driven Mechanisms Between Phosphorylated Caveolin-1 and Contractile Actin Assemblies Instruct Persistent Cell Migration

doi: 10.3389/fcell.2021.665919

Figure Lengend Snippet: Actin protrusive network is upregulated by AMPK-Rac1-PAK1-Cofilin signaling cascade in cells lacking CAV-1. (A) Immunofluorescence microscopy analysis demonstrating that more pronounced endogenous ARPC2 colocalizes with F-actin (visualized by Alexa 568 phalloidin) on the lamellipodia protrusions in CAV-1 deficient cells. Magnified regions of cell edges on the right show the distribution of ARPC2 in WT and CAV-1 KO cells. Bars, 10 μm (in cell images) and 5 μm (in the magnified box). Quantification of width of lamellipodial protrusions are shown on the right. n = 16 regions from 16 cells for each group. (B) Representative images of membrane ruffling by time-lapse microscope. 1-pixel-wide areas were cut out to generate a 300 frame 2 s interval kymograph. Yellow dashed lines indicate the track of cell movement. An enlarged region is displayed on the right, vertical dashed lines show the membrane protrusion distance, while horizontal dashed lines mark the duration of protrusion. Bars, 10 μm (in cell images) and 2 μm (in the magnified box). Quantification of protrusion rate are shown on the right. n = 16 regions from 16 cells for each group. (C) P-AMPK (Thr172) and total AMPK were detected from the lysates of each group by western blotting. Please note that CAV-1(Y14F)-mEGFP can’t be detected by using phospho-CAV-1(Tyr14) antibody. Asterisk denotes the non-specific band. Quantification of P-AMPK (Thr172) levels (compared to total AMPK) from each group was shown on the right panel. n = 3. (D) Pull-down assays were performed for WT, CAV-1 KO, and CAV-1 KO; CAV-1-mEGFP re-expressed cells. Proteins bound to GST-PAK binding domain were analyzed by western blots and further quantified (compared to total Rac1) based on the band’s intensity. n = 3. (E) Western blot analysis and quantification (compared to total PAK1 and Cofilin) of the levels of phosphorylated PAK1 (Thr423) and Cofilin (Ser3) in WT and CAV-1 KO cell lysates. n = 3. (F) Immunostaining and quantification of endogenous P-Cofilin (Ser3) and F-actin distribution in WT and CAV-1 KO cells. A 16 μm length line was used to generate a line profile to illustrate the co-localization of P-Cofilin (Ser3) and F-actin. The lamellipodia region was enlarged on the right, and 2 μm width region was chosen to analyze the mean intensity of P-Cofilin (Ser3) on the leading edge. Bars, 10 μm (in cell images) and 5 μm (in the magnified box). n = 18 regions from 18 cells for each group. (G) Immunostaining and quantification of endogenous P-cofilin (Ser3) distribution upon compound C treatment in WT and CAV-1 KO cells. Magnified regions represent the lamellipodia region. Bars, 10 μm (in cell images) and 5 μm (in the magnified box). n = 18 regions from 18 cells for each group. (H) Western blot analysis and quantification of the phosphorylated levels of AMPK (Thr172), PAK1 (Thr423), Cofilin (Ser3) and activity of Rac1 and RhoA upon compound C treatment. n = 3. In panel (C–E,H) , the obtained intensity value from wild-type cells was set to 1. All the data are presented as mean ± SD. In (C,G,H) , *** P < 0.001; * P < 0.05; N.A., not significant (one-way ANOVA). In (D–F) , *** P < 0.001; ** P < 0.01 (unpaired t -test). All the data are from three independent experiments.

Article Snippet: The following antibodies were used in this study: CAV-1 (D46G3) rabbit antibody (1:1,000 dilution for WB, 1:200 for IF; #3267, Cell Signaling, Beverly, MO, United States); Phospho-CAV-1 (Tyr14) rabbit antibody (dilution 1:1,000 for WB; #3251, Cell signaling); AMPK rabbit antibody (dilution 1:500 for WB; #SAB4502329, Sigma, St. Louis, MO, United States); P-AMPK (Thr172) rabbit antibody (dilution 1:500 for WB, 1:100 for IF; #2531S, Cell Signaling); Cofilin (E-8) mouse antibody (dilution 1:1,000 for WB; #sc-376476, Santa Cruz, Dallas, TX, United States); Phospho-Cofilin (Ser3) rabbit antibody (dilution 1:1,000 for WB, 1:200 for IF; #3313, Cell signaling); p190RhoGAP rabbit antibody (dilution 1:2,000 for WB; #26789, Proteintech, Rosemont, IL, United States); FAK rabbit antibody (dilution 1:1,000 for WB; #3285, Cell Signaling); Phospho-FAK (Tyr397) rabbit antibody (dilution 1:1,000 for WB; #3283, Cell Signaling); PAK1 rabbit antibody (dilution 1:1,000; #2602, Cell Signaling); Phospho-PAK1 (Thr423)/PAK2 (Thr402) rabbit antibody (dilution 1:1,000 for WB, 1:200 for IF; #2601, Cell Signaling); Tpm4.2 (LC24) mouse antibody (dilution 1:500 for WB and IF; a kind gift from Peter W. Gunning, UNSW Australia); Phospho-myosin light chain 2 (Thr18/Ser19) rabbit antibody (dilution 1:500 for WB, 1:200 for IF; #3674, Cell Signaling); Myosin light chain mouse antibody (dilution 1:1,000 for WB; #M4401, Sigma); Vinculin mouse antibody (dilution 1:100 for IF; #V9131, Sigma); ARPC2 rabbit antibody (dilution 1:1,000 for WB and IF; #15058, Proteintech, Rosemont, IL, United States); Myosin-18B rabbit antibody (dilution 1:500 for WB; #HPA000953, Sigma); Rab8 rabbit antibody (dilution 1:100 for IF; #R5530, Sigma); and GAPDH mouse polyclonal antibody (dilution 1:1,000 for WB; #G8795, Sigma).

Techniques: Immunofluorescence, Microscopy, Membrane, Western Blot, Binding Assay, Immunostaining, Activity Assay