Journal: International immunopharmacology

Article Title: RACGAP1 is a pivotal gene in lung adenocarcinoma-associated membranous nephropathy: Based on comprehensive bioinformatics analysis and machine learning.

doi: 10.1016/j.intimp.2024.112783

Figure Lengend Snippet: Fig. 3. LASSO algorithm identified 4 significant genes (A). The RFB algorithm confirmed 22 genes (B). The SVM algorithm identified 4 genes (C). Venn diagram of 4 hub genes of LUAD-associated MN (D). ROC curve showed four genes exhibited satisfactory efficacy in discriminating MN (COL3A1: 0.994, PSENEN: 0.994, RACGAP1: 0.970, TNFRSF10B: 0.964) (E).

Article Snippet: The following antibodies (1) COL3A1 antibody (68320–1-Ig, Proteintech) diluted 1:200, (2) PSENEN antibody (ab154830, Abcam, Cambridge) diluted 1:100, (3) RACGAP1 antibody (13739–1-AP, Proteintech) diluted 1:200, (4) TNFRSF10B antibody (ab230969, Abcam, Cambridge) diluted 1:1000 were incubated overnight at 4◦ C, washed with phosphate buffer saline (PBS) obtained from Thermo Fisher Scientific, and then incubated at 4◦ C for 2 h. C, washed with PBS and stained with horseradish peroxidase and diaminobenzidine to visualize the reaction.

Techniques:

Journal: International immunopharmacology

Article Title: RACGAP1 is a pivotal gene in lung adenocarcinoma-associated membranous nephropathy: Based on comprehensive bioinformatics analysis and machine learning.

doi: 10.1016/j.intimp.2024.112783

Figure Lengend Snippet: Fig. 4. Correlation analysis of 4 genes (A). Survival analysis showed a worse prognosis with higher expression of gene COL3A1 (B), PSENEN (C), RACGAP1 (D), and TNFRSF10B (E) in the TCGA-LUAD cohort.

Article Snippet: The following antibodies (1) COL3A1 antibody (68320–1-Ig, Proteintech) diluted 1:200, (2) PSENEN antibody (ab154830, Abcam, Cambridge) diluted 1:100, (3) RACGAP1 antibody (13739–1-AP, Proteintech) diluted 1:200, (4) TNFRSF10B antibody (ab230969, Abcam, Cambridge) diluted 1:1000 were incubated overnight at 4◦ C, washed with phosphate buffer saline (PBS) obtained from Thermo Fisher Scientific, and then incubated at 4◦ C for 2 h. C, washed with PBS and stained with horseradish peroxidase and diaminobenzidine to visualize the reaction.

Techniques: Expressing

Journal: International immunopharmacology

Article Title: RACGAP1 is a pivotal gene in lung adenocarcinoma-associated membranous nephropathy: Based on comprehensive bioinformatics analysis and machine learning.

doi: 10.1016/j.intimp.2024.112783

Figure Lengend Snippet: Fig. 5. Biological functions of LUAD-associated MN genes analyzed by the STRING database. COL3A1 interacts with 10 proteins (A) in cell adhesion, skin collagen fiber formation, osteogenic amino acid metabolism, and extracellular fiber structure process (B, C). PSENEN interacts with 11 proteins (D) in amyloid protein metabolism, Notch signaling pathway, and membrane protein hydrolysis process (E, F). RACGAP1 interacts with 10 proteins (G) in cell division, tight junctions, and phagocytosis (H, I) process. TNFRSF10B interacts with 10 proteins (J) in apoptosis, TNF signaling pathway, cell death, TNF signaling pathway, and NK-kappaB signaling pathway (K, L).

Article Snippet: The following antibodies (1) COL3A1 antibody (68320–1-Ig, Proteintech) diluted 1:200, (2) PSENEN antibody (ab154830, Abcam, Cambridge) diluted 1:100, (3) RACGAP1 antibody (13739–1-AP, Proteintech) diluted 1:200, (4) TNFRSF10B antibody (ab230969, Abcam, Cambridge) diluted 1:1000 were incubated overnight at 4◦ C, washed with phosphate buffer saline (PBS) obtained from Thermo Fisher Scientific, and then incubated at 4◦ C for 2 h. C, washed with PBS and stained with horseradish peroxidase and diaminobenzidine to visualize the reaction.

Techniques: Membrane

Journal: International immunopharmacology

Article Title: RACGAP1 is a pivotal gene in lung adenocarcinoma-associated membranous nephropathy: Based on comprehensive bioinformatics analysis and machine learning.

doi: 10.1016/j.intimp.2024.112783

Figure Lengend Snippet: Fig. 6. Comparative Toxicogenomics Database (CTD) analysis. COL3A1 associates with proteinuria, adenocarcinoma, and focal segmental glomerulosclerosis (A). PSENEN associates with adenocarcinoma, proteinuria, and focal segmental glomerulosclerosis (B). RACGAP1 associates with adenocarcinoma, proteinuria, and nephrotic syndrome (C). TNFRSF10B associates with adenocarcinoma, non-small-cell lung cancer, and proteinuria (D).

Article Snippet: The following antibodies (1) COL3A1 antibody (68320–1-Ig, Proteintech) diluted 1:200, (2) PSENEN antibody (ab154830, Abcam, Cambridge) diluted 1:100, (3) RACGAP1 antibody (13739–1-AP, Proteintech) diluted 1:200, (4) TNFRSF10B antibody (ab230969, Abcam, Cambridge) diluted 1:1000 were incubated overnight at 4◦ C, washed with phosphate buffer saline (PBS) obtained from Thermo Fisher Scientific, and then incubated at 4◦ C for 2 h. C, washed with PBS and stained with horseradish peroxidase and diaminobenzidine to visualize the reaction.

Techniques:

Journal: International immunopharmacology

Article Title: RACGAP1 is a pivotal gene in lung adenocarcinoma-associated membranous nephropathy: Based on comprehensive bioinformatics analysis and machine learning.

doi: 10.1016/j.intimp.2024.112783

Figure Lengend Snippet: Fig. 7. Immunohistochemistry showed a significant difference in the expression of RACGAP1(A, B, C), COL3A1(D, E, F), and PSENEN(G, H, I) between PMN and LUAD-associated MN glomeruli. RACGAP1 was especially expressed on the glomerular basement membrane in LUAD-associated MN(B). TNFRSF10B expression wasn’t different between the two groups (J, K, L) (original magnification × 400 scale: 50um).

Article Snippet: The following antibodies (1) COL3A1 antibody (68320–1-Ig, Proteintech) diluted 1:200, (2) PSENEN antibody (ab154830, Abcam, Cambridge) diluted 1:100, (3) RACGAP1 antibody (13739–1-AP, Proteintech) diluted 1:200, (4) TNFRSF10B antibody (ab230969, Abcam, Cambridge) diluted 1:1000 were incubated overnight at 4◦ C, washed with phosphate buffer saline (PBS) obtained from Thermo Fisher Scientific, and then incubated at 4◦ C for 2 h. C, washed with PBS and stained with horseradish peroxidase and diaminobenzidine to visualize the reaction.

Techniques: Immunohistochemistry, Expressing, Membrane

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: RACGAP1 protein expression in lung cancer tissues and normal tissues. ( a ) Immunohistochemical staining of RACGAP1 in LUAD tissues and normal lung tissues from the HPA database. ( b ) RACGAP1 protein expression in LUAD tissues and normal lung tissues from CPTAC samples. ( c ) RACGAP1 protein levels were determined via IHC using tissue microarrays in 75 pairs of lung adenocarcinoma and adjacent noncancerous lung tissues. The lower right panels show representative images captured at × 4 and × 20 magnification. Each datapoint in the plot represents one tissue sample. Error bars represent the median ± range. T, lung cancer tissue; N, noncancerous lung tissue. * P < 0.05 vs. normal tissues.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: Analysis of RACGAP1 alterations in LUAD. ( a ) CNA and mutation frequency data for RACGAP1 in LUAD were analyzed through the cBioPortal website. ( b ) OncoPrint visual summary of alterations on a query of RACGAP1. Variations in frequency included missense mutation (green), truncating mutation (gray), amplification (red), and deep deletions (blue).

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Mutagenesis, Amplification

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: Correlation between RACGAP1 expression and clinical features of LUAD patients. Correlation between RACGAP1 expression and ( a ) age, ( b ) sex, ( c ) smoking habits, ( d ) stage, ( e ) nodal metastasis, or ( f ) TP53 mutation status of LUAD patients.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Expressing, Mutagenesis

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: Correlation between RACGAP1 expression and survival of LUAD patients. ( a ) The correlation between RACGAP1 expression and OS of LUAD patients was analyzed using GEPIA. ( b – d ) The correlation between RACGAP1 expression and OS, PFS, or PPS of LUAD patients was analyzed through the Kaplan–Meier Plotter. OS, overall survival; PFS, free-progression survival; PPS, post-progression survival.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: RACGAP1 knockdown inhibited cell growth in A549 cells. A549 cells were transfected with si-Ctrl or three independent siRNAs for si-RACGAP1. ( a ) Protein levels of RACGAP1 were determined using Western blot analysis. Representative images of RACGAP1 and β-actin are shown and quantitated using ImageLab software. β-actin was used as an internal control. * P < 0.05 vs. si-Ctrl. ( b ) Cell viability was determined using the CCK-8 assay. Data were normalized to the viability on day 1 and are represented as fold change. * P < 0.05 vs. si-Ctrl. ( c ) The colony formation assay was used to determine cell survival. After obtaining the images, colony formation was calculated and normalized to the survival of control cells. * P < 0.05 vs. si-Ctrl.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Knockdown, Transfection, Western Blot, Software, Control, CCK-8 Assay, Colony Assay

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: RACGAP1 knockdown inhibited cell proliferation in A549 cells. A549 cells were transfected with si-Ctrl or three independent siRNAs for si-RACGAP1. ( a ) Cell cycle progression was determined through PI staining and FACS analysis. Representative plots and percentage of cells at different stages (G0/G1, G2/M, and S phases) were presented. * P < 0.05 vs. si-Ctrl. ( b ) Protein levels of CDK4 and CyclinD1 after RACGAP1 knockdown were determined using Western blot analysis. Representative images are shown and quantitated using ImageLab software. β-actin was used as an internal control. * P < 0.05 vs. si-Ctrl.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Knockdown, Transfection, Staining, Western Blot, Software, Control

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: RACGAP1 knockdown induced cell apoptosis in A549 cells. A549 cells were transfected with si-Ctrl or three independent siRNAs for si-RACGAP1. ( a ) Cell apoptosis was determined using Annexin V/PI staining and FACS analysis. Representative plots and percentage of apoptotic cells are presented. *P < 0.05 vs. si-Ctrl. ( b ) Protein levels of Bax and Bcl-2 after RACGAP1 knockdown were determined using Western blot analysis. Representative images are shown and quantitated using ImageLab software. β-actin was used as an internal control. * P < 0.05 vs. si-Ctrl.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Knockdown, Transfection, Staining, Western Blot, Software, Control

Journal: Scientific Reports

Article Title: RACGAP1 promotes lung cancer cell proliferation through the PI3K/AKT signaling pathway

doi: 10.1038/s41598-024-58539-0

Figure Lengend Snippet: RACGAP1 knockdown inhibited the PI3K/AKT signaling pathway in A549 cells. A549 cells were transfected with si-Ctrl or si-RACGAP1. Protein levels of PI3K, p-AKT, and AKT after RACGAP1 knockdown were determined using Western blot analysis. Representative images are shown and quantitated using ImageLab software. GAPDH was used as an internal control. * P < 0.05 vs. si-Ctrl.

Article Snippet: The antibody against RACGAP1 (13739-1-AP) was purchased from Proteintech Group (Wuhan, Hubei, China).

Techniques: Knockdown, Transfection, Western Blot, Software, Control