Journal: Cell Genomics

Article Title: Variant-to-function analysis of the childhood obesity chr12q13 locus implicates rs7132908 as a causal variant within the 3′ UTR of FAIM2

doi: 10.1016/j.xgen.2024.100556

Figure Lengend Snippet: rs7132908 regulates FAIM2 expression with allele and cell-type specificity (A) Chromatin accessibility represented by ATAC-seq tracks depicting normalized reads and chromatin loops at the TAD containing rs7132908 in neural cell types. Chromatin loops represent significant contacts between regions of open chromatin that harbored rs7132908 and a gene promoter. Gray dashed vertical line indicates rs7132908 position. (B) Graphic representation of firefly luciferase reporter vectors used in luciferase reporter assays. (C‒F) Fold change of firefly luciferase fluorescence normalized to the promoter only control vector driven by the FAIM2 promoter in primary astrocytes ( n = 7 biological replicates) (C), FAIM2 promoter in HEK293Ts ( n = 7 biological replicates) (D), LIMA1 promoter in primary astrocytes ( n = 8 biological replicates) (E), and RACGAP1 promoter in primary astrocytes ( n = 9 biological replicates) (F). Data are represented as mean ± SD. ∗ p value <0.05, ∗∗ p value <0.01, ∗∗∗ p value <0.001 by one-way ANOVA with Tukey’s correction for multiple comparisons.

Article Snippet: To generate DNA fragments containing the FAIM2 , LIMA1 , and RACGAP1 promoter sequences, we also designed PCR primers ( ) to amplify these regions and used promoter clones (purchased from GeneCopoeia) as PCR templates.

Techniques: Expressing, Luciferase, Fluorescence, Control, Plasmid Preparation

Journal: Cell Genomics

Article Title: Variant-to-function analysis of the childhood obesity chr12q13 locus implicates rs7132908 as a causal variant within the 3′ UTR of FAIM2

doi: 10.1016/j.xgen.2024.100556

Figure Lengend Snippet:

Article Snippet: To generate DNA fragments containing the FAIM2 , LIMA1 , and RACGAP1 promoter sequences, we also designed PCR primers ( ) to amplify these regions and used promoter clones (purchased from GeneCopoeia) as PCR templates.

Techniques: Recombinant, Saline, Modification, Transfection, Electron Microscopy, Membrane, Knock-Out, Gel Extraction, Plasmid Preparation, Mutagenesis, Reporter Assay, Sensitive Assay, Gene Expression, Purification, Library Quantification, Software, Lysis, Cell Culture

Journal: Oncotarget

Article Title: Comparative evaluation of three proliferation markers, Ki-67, TOP2A, and RacGAP1, in bronchopulmonary neuroendocrine neoplasms: Issues and prospects

doi: 10.18632/oncotarget.9747

Figure Lengend Snippet: Overview of the patient population in terms of the investigated markers in the different evaluation methods. BP-NEN – bronchopulmonary neuroendocrine neoplasms; TC – typical carcinoids; AC – atypical carcinoids; SCLC – small cell lung cancer; LCNEC – large cell neuroendocrine lung carcinoma; FFPE – formalin-fixed, paraffin-embedded; TOP2A – Topoisomerase 2 alpha; RacGAP1 – Rac GTPase activating protein 1; IHC – immunohistochemistry.

Article Snippet: After dewaxing and microwaving of the sections in 10 mM citrate buffer (pH = 6.0) for 16 minutes at 600 W, the slices were incubated overnight at 4°C with the primary antibodies (Ki-67 (mouse monoclonal, 1:50, Dako Germany GmbH, Hamburg, GER), TOP2A (rabbit monoclonal, 1:100, Abcam, Cambridge, UK), RacGAP1 (rabbit polyclonal, 1:350, Atlas Antibodies, Stockholm, SWE)).

Techniques: Formalin-fixed Paraffin-Embedded, Immunohistochemistry

Journal: Oncotarget

Article Title: Comparative evaluation of three proliferation markers, Ki-67, TOP2A, and RacGAP1, in bronchopulmonary neuroendocrine neoplasms: Issues and prospects

doi: 10.18632/oncotarget.9747

Figure Lengend Snippet: The boxplots are depicting the TOP2A and RacGAP1 protein (left) and mRNA (right) levels in the different BP-NEN entities as evaluated by immunohistochemistry and RT-qPCR. To the right the respective Kaplan-Meier-Analyses of each marker in the IHC investigation are shown. Blue indicates low, green moderate and red high TOP2A/RacGAP1 expression levels, according to the cut-off limits of the ROC analysis between TC – AC and AC – SCLC. With both markers, high expression levels imply poor patient survival.

Article Snippet: After dewaxing and microwaving of the sections in 10 mM citrate buffer (pH = 6.0) for 16 minutes at 600 W, the slices were incubated overnight at 4°C with the primary antibodies (Ki-67 (mouse monoclonal, 1:50, Dako Germany GmbH, Hamburg, GER), TOP2A (rabbit monoclonal, 1:100, Abcam, Cambridge, UK), RacGAP1 (rabbit polyclonal, 1:350, Atlas Antibodies, Stockholm, SWE)).

Techniques: Immunohistochemistry, Quantitative RT-PCR, Marker, Expressing

Journal: Oncotarget

Article Title: Comparative evaluation of three proliferation markers, Ki-67, TOP2A, and RacGAP1, in bronchopulmonary neuroendocrine neoplasms: Issues and prospects

doi: 10.18632/oncotarget.9747

Figure Lengend Snippet: Spearman's rank correlations of Ki-67, TOP2A and RacGAP1 protein and mRNA levels with clinical data and extent of necrosis

Article Snippet: After dewaxing and microwaving of the sections in 10 mM citrate buffer (pH = 6.0) for 16 minutes at 600 W, the slices were incubated overnight at 4°C with the primary antibodies (Ki-67 (mouse monoclonal, 1:50, Dako Germany GmbH, Hamburg, GER), TOP2A (rabbit monoclonal, 1:100, Abcam, Cambridge, UK), RacGAP1 (rabbit polyclonal, 1:350, Atlas Antibodies, Stockholm, SWE)).

Techniques:

Journal: Oncotarget

Article Title: Comparative evaluation of three proliferation markers, Ki-67, TOP2A, and RacGAP1, in bronchopulmonary neuroendocrine neoplasms: Issues and prospects

doi: 10.18632/oncotarget.9747

Figure Lengend Snippet: Below two examples for a strong positivity of RacGAP1 protein expression at the invasive front are shown (magnification 10 × 10). The graph above depicts a Kaplan-Meier-Analysis with a high RacGAP1 protein expression (red line) and a weak/no RacGAP1 expression (blue line) at the invasive front. According to the Log-Rank Test the curves differ highly significantly: High RacGAP1 expression at the invasive front indicates poor patient prognosis.

Article Snippet: After dewaxing and microwaving of the sections in 10 mM citrate buffer (pH = 6.0) for 16 minutes at 600 W, the slices were incubated overnight at 4°C with the primary antibodies (Ki-67 (mouse monoclonal, 1:50, Dako Germany GmbH, Hamburg, GER), TOP2A (rabbit monoclonal, 1:100, Abcam, Cambridge, UK), RacGAP1 (rabbit polyclonal, 1:350, Atlas Antibodies, Stockholm, SWE)).

Techniques: Expressing

Journal: Oncotarget

Article Title: Comparative evaluation of three proliferation markers, Ki-67, TOP2A, and RacGAP1, in bronchopulmonary neuroendocrine neoplasms: Issues and prospects

doi: 10.18632/oncotarget.9747

Figure Lengend Snippet: Immunohistochemical staining of Ki-67, TOP2A and RacGAP1 at the same location of each one TC, AC, SCLC and LCNEC sample (magnification 40 × 10).

Article Snippet: After dewaxing and microwaving of the sections in 10 mM citrate buffer (pH = 6.0) for 16 minutes at 600 W, the slices were incubated overnight at 4°C with the primary antibodies (Ki-67 (mouse monoclonal, 1:50, Dako Germany GmbH, Hamburg, GER), TOP2A (rabbit monoclonal, 1:100, Abcam, Cambridge, UK), RacGAP1 (rabbit polyclonal, 1:350, Atlas Antibodies, Stockholm, SWE)).

Techniques: Immunohistochemical staining, Staining

Journal: Cancers

Article Title: Lambda-Carrageenan Enhances the Effects of Radiation Therapy in Cancer Treatment by Suppressing Cancer Cell Invasion and Metastasis through Racgap1 Inhibition

doi: 10.3390/cancers11081192

Figure Lengend Snippet: Upregulation of RacGAP1 is involved in cancer cell survival and invasion after IR. ( A ) Gene expression was analyzed by cDNA microarray. RacGAP1 expression level is shown by the heat map. The values were normalized to the untreated group. ( B ) RacGAP1 protein expressions with different doses of IR were analyzed in MDA-MB-231, FaDu, PANC-1, and 4T1 cell lines. ( C ) MDA-MB-231 cells were transfected with siRNA duplexes targeting RacGAP1 (# 1 or # 2) or the control sequence, as indicated. Cell lysates were subjected to western blot two days after transfection. ( D – H ) MDA-MB-231 cells were transfected with siRNAs and incubated for two days, and then treated with 4 Gy IR. Each experiment was performed 24 h after IR treatment. Cell viability was quantified by cell counting ( D ). Apoptotic cells were measured by Annexin V-FITC staining and flow cytometry ( E ). ROS was measured by DCFDA ( F ). Cells were treated with staining solution and PI, and then the cell cycle was analyzed by flow cytometry. The percentage of polyploid cells in each group was normalized with control group ( G ). The invasive activity was measured by Matrigel chemoinvasion assay ( H ). Columns, mean ( n ≥ 3); bars, SE. *, p < 0.05; **, p < 0.01. Data shown in ( D – H ) were normalized to that of MDA-MB-231 cells transfected with siRNA control. For the continuous full length-image of Western blot signals, please refer to .

Article Snippet: Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight.

Techniques: Gene Expression, Microarray, Expressing, Transfection, Control, Sequencing, Western Blot, Incubation, Cell Counting, Staining, Flow Cytometry, Activity Assay

Journal: Cancers

Article Title: Lambda-Carrageenan Enhances the Effects of Radiation Therapy in Cancer Treatment by Suppressing Cancer Cell Invasion and Metastasis through Racgap1 Inhibition

doi: 10.3390/cancers11081192

Figure Lengend Snippet: RacGAP1 expression is suppressed by CGN in MDA-MB-231 cells. ( A ) Protein expression of RacGAP1 was analyzed after treatment with different concentrations of CGN in MDA-MB-231 cells. Total cell lysates were subjected to western blot. ( B ) RacGAP1 protein expression was analyzed using cell lysates after IR and/or CGN treatment in MDA-MB-231 cells. ( C ) Immunofluorescence images show RacGAP1 (green), α-tubulin (red) and nuclei (blue). Bar, 25 μm. ( D ) RacGAP1-mVenus expression was induced by doxycycline in MDA-MB-231 cells. RacGAP1 expression was analyzed by western blot using anti-RacGAP1 antibody. Dox, doxycycline. ( E ) Cell number was quantified by cell counting. Columns, mean ( n = 3); bars, SE. *, p < 0.05; **, p < 0.01. For the continuous full length-image of Western blot signals, please refer to .

Article Snippet: Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight.

Techniques: Expressing, Western Blot, Immunofluorescence, Cell Counting

Journal: Cancers

Article Title: Lambda-Carrageenan Enhances the Effects of Radiation Therapy in Cancer Treatment by Suppressing Cancer Cell Invasion and Metastasis through Racgap1 Inhibition

doi: 10.3390/cancers11081192

Figure Lengend Snippet: Radiation followed by CGN treatment decreases tumor size and metastasis in a 4T1 mouse xenograft model. ( A ) Treatment schedule of each group. Mouse 4T1 cells were injected into Balb/c mice ( n = 6–7 in each group). ( B ) 4T1 tumor sizes were measured and normalized to size at day 12 in each group. ( C ) Relative tumor size for each group was measured on Day 25. Scatter plot; mean ± SE. *, p < 0.05. ( D ) Representative H&E images of lung sections. Bar, 500 µm. Arrow, metastatic lung nodules. The number of metastatic lung nodules was counted (right panel). Scatter plot; mean ± SE. *, p < 0.05. ( E ) Sections from 4T1 tumors were subjected to IHC staining with antibodies against RacGAP1. Bar, 100 µm. RacGAP1 expression level was determined by scoring, as described in the methods (right panel). Scatter plot; bars, mean ( n = 5). *, p < 0.05.

Article Snippet: Each slide was incubated in 2% blocking buffer (Roche, Basel, Switzerland) for 1 h, and then incubated with a RacGAP1 primary antibody (Proteintech) overnight.

Techniques: Injection, Immunohistochemistry, Expressing

Journal: BMC cancer

Article Title: RACGAP1 promotes the progression and poor prognosis of lung adenocarcinoma through its effects on the cell cycle and tumor stemness.

doi: 10.1186/s12885-023-11761-x

Figure Lengend Snippet: Fig. 5 Flow cytometry analysis of cell cycle and WB validation of cell cycle protein and pluripotency-related protein expression. (A) GSEA analysis was performed on RACGAP1; (B-C) Flow cytometry analysis of cell cycle was conducted on A549 and H1975 cells after transfection with si-RACGAP1; (D) Compared to the negative control (NC), specific siRNA targeting RACGAP1 successfully inhibited the expression of RACGAP1 protein as well as cell cycle-related proteins CDK2, CDK4 and CDK1; (E) Western blot results showed that the expression of pluripotency-related markers NIFK and NANOG was downregulated after transfection with si-RACGAP1 (The blot has been cropped and the original blot is shown in the supplementary file.) (***p < 0.001; **p < 0.01; *p < 0.05; ns: not significant)

Article Snippet: Antibodies against RACGAP1 (71KDa, 1:500), CDK2 (35KDa, 1:1000), CDK4 (34KDa, 1:1000), CDK1 (34KDa, 1:1000), NIFK (KI-67) (39KDa, 1:1000) and ACTB (42KDa, 1:1000) were obtained from ABclonal.

Techniques: Flow Cytometry, Biomarker Discovery, Expressing, Transfection, Negative Control, Western Blot

Journal: Cell death & disease

Article Title: PLAGL2 promotes bladder cancer progression via RACGAP1/RhoA GTPase/YAP1 signaling.

doi: 10.1038/s41419-023-05970-2

Figure Lengend Snippet: Fig. 5 PLAGL2 transcriptionally activates RACGAP1 to repress Hippo signaling. Immunofluorescence (A) and immunoblotting assays (B) indicated the expression of RACGAP1 in the indicated cells after PLAGL2 interference. C Schematic representation of the RACGAP1 promoter. The sequences of wild-type/mutant PLAGL2 binding sites are indicated. D Luciferase activity assays were performed in the indicated cells transfected with wild-type (RACGAP1-WT) or mutant-type (RACGAP1-mut) RACGAP1 promoter-reporter plasmids. E ChIP‒qPCR analysis displays RACGAP1 enrichment on the promoter of RACGAP1. IgG indicates the negative control. F ChIP assays were employed in the indicated cells using anti-p300, anti-RNA polymerase II, and anti-H3K4me3 antibodies. *P < 0.05, **P < 0.01, ***P < 0.001, n.s no significance.

Article Snippet: IHC was first incubated with primary antibodies against PLAGL2 (#11540- 1-AP; Proteintech), RACGAP1 (#NBP1-33455; Novus), YAP1 (#12395; Cell Signaling Technology), and Ki-67 (#27309-1-AP; Proteintech) at 4 °C overnight.

Techniques: Western Blot, Expressing, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Negative Control

Journal: Cell death & disease

Article Title: PLAGL2 promotes bladder cancer progression via RACGAP1/RhoA GTPase/YAP1 signaling.

doi: 10.1038/s41419-023-05970-2

Figure Lengend Snippet: Fig. 6 PLAGL2 promotes YAP1 signaling by activating RhoA. A Immunoblotting assays of the indicated protein expression in T24 and RT4 cells after PLAGL2, RACGAP1, and simvastatin (5 μM, 24 h) interference. GAPDH and P84 were used as loading controls for total and nuclear extracts, respectively. B Immunofluorescence assays indicated the expression and subcellular localization of YAP1 in T24 and RT4 cells after PLAGL2 interference and simvastatin treatment. The percentage of subcellular localization of YAP1 is shown. N nucleus, C cytoplasm. C HOP/ HIP luciferase reporter assays showed YAP1/TAZ-TEAD transcriptional activity in T24 and RT4 cells after PLAGL2 interference and simvastatin/ verteporfin treatment. D The mRNA levels of YAP1-targeted genes in T24 and RT4 cells after PLAGL2 interference and simvastatin/verteporfin treatment. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: IHC was first incubated with primary antibodies against PLAGL2 (#11540- 1-AP; Proteintech), RACGAP1 (#NBP1-33455; Novus), YAP1 (#12395; Cell Signaling Technology), and Ki-67 (#27309-1-AP; Proteintech) at 4 °C overnight.

Techniques: Western Blot, Expressing, Luciferase, Activity Assay

Journal: Cell death & disease

Article Title: PLAGL2 promotes bladder cancer progression via RACGAP1/RhoA GTPase/YAP1 signaling.

doi: 10.1038/s41419-023-05970-2

Figure Lengend Snippet: Fig. 8 Clinical relevance of the PLAGL2/RACGAP1/YAP1 axis in human BCa. A Representative IHC images of PLAGL2, RACGAP1, YAP1, and p-YAP1 in 164 BCa specimens. B Upper panel: The proportion of specimens with the indicated RACGAP1 between those with high and low PLAGL2 expression. Bottom panel: The proportion of specimens with the indicated nuclear YAP1 between those with high and low PLAGL2 expression. C DFS of BCa patients with the indicated expression of proteins. D The schematic diagram illustrates that PLAGL2 promotes BCa progression via RACGAP1-mediated RhoA activation. ***P < 0.001.

Article Snippet: IHC was first incubated with primary antibodies against PLAGL2 (#11540- 1-AP; Proteintech), RACGAP1 (#NBP1-33455; Novus), YAP1 (#12395; Cell Signaling Technology), and Ki-67 (#27309-1-AP; Proteintech) at 4 °C overnight.

Techniques: Expressing, Activation Assay

Journal: Frontiers in Immunology

Article Title: Exploratory transcriptomic analysis of mouse articular cartilage in response to tissue inhibitor of metalloproteinase 3 identifies inflammation-associated gene expression changes

doi: 10.3389/fimmu.2026.1794078

Figure Lengend Snippet: RNA-seq analysis of TIMP-3–treated cartilage under hypoxia. (A) MA plot showing differential expression between TIMP-3–treated and control articular cartilage, with log 2 FC plotted against average log 2 CPM expression (n = 4). Grey: no change, red: upregulated, blue: downregulated (P < 0.01, |log 2 FC| > 0.58). The 10 most highly expressed regulated genes are labelled. G1 and G2 correspond to E430024I08Rik and AC127578.1 respectively. Complete gene lists are in . (B) Heatmap of differentially expressed genes (log 2 FC versus the mean of the controls). Genes are ordered by hierarchical clustering. Sample labels: HC1-4: hypoxia controls; HT1-4: hypoxia TIMP-3-treated. (C) Protein–protein interaction (PPI) network corresponding to genes downregulated by TIMP-3 under hypoxia (P <0.01 by quasi-likelihood F-test in edgeR, |log 2 FC| > 0.58), generated using STRING (default interaction score ≥ 0.400). All nodes represent the initially filtered gene list and are included to show the network context and highlight that only Pbk and Racgap1 display a documented interaction. Line colors indicate evidence type: green, text mining; pink, experimental; black, co-expression. Combined interaction score: 0.711. (D) RT-qPCR validation. RNA-seq (left) and RT-qPCR (right) data shown as log 2 FC versus the mean of the controls (mean ± SD, n = 4). RNA-seq: log 2 FC were calculated from normalized CPM values; § FDR < 0.05 (hypoxia effect), # P < 0.01 (TIMP-3 effect) by quasi-likelihood F-test applied to raw counts (edgeR); RT-qPCR: *P < 0.05, **P < 0.01 by unpaired two-sided Welch’s t-test on log 2 FC values (-ΔΔCq). CPM, counts per million; Ctrl, control.

Article Snippet: TaqMan assays used included Saa3 (Mm00441203_m1), Il17b (Mm01258783_m1), Mmp3 (Mm00440295_m1), Lcn2 (Mm01324470_m1), Pbk (Mm01313789_g1), Racgap1 (Mm00488845_m1), Hprt (Mm00446968_m1).

Techniques: RNA Sequencing, Quantitative Proteomics, Control, Expressing, Generated, Quantitative RT-PCR, Biomarker Discovery

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Expression of Rac GTPase activating protein 1 (RacGAP1) in epithelial ovarian cancer (EOC) tissues and its correlations with clinicopathological parameters. A, Representative high expression of RacGAP1 in the nucleus of EOC cells. B, Representative low expression of RacGAP1 in EOC tissue samples. Proportion of high‐RacGAP1 expression is higher in patients with (C) poor differentiation grade, (D) positive lymph node metastasis, (E) advanced International Federation of Gynecology and Obstetrics (FIGO) stage, and (F) positive tumor cells in ascetic fluid. Data were analyzed by χ 2 ‐test

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Expressing

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Kaplan‐Meier curves for overall survival and progression‐free survival according to Rac GTPase activating protein 1 (RacGAP1) expression and different clinicopathological features in epithelial ovarian cancer (EOC) patients. Patients with (A) higher RacGAP1 expression level, (B) positive lymph node metastasis, (C) advanced International Federation of Gynecology and Obstetrics (FIGO) stage and (D) higher serum CA‐125 level exhibited unfavorable overall survival. E‐H, Similarly, progression‐free survival was also evaluated by plotting Kaplan‐Meier curves. All statistical significance was compared by using log‐rank test

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Expressing

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Role of Rac GTPase activating protein 1 (RacGAP1) in promoting epithelial ovarian cancer (EOC) cell migration and invasion. A, RNAi efficiency of RacGAP1‐siRNAs was tested by Western blotting, using scrambled siRNA (scram) as the negative control. B, Migration and (C) invasion capacities of OV‐90 cells were upregulated upon RacGAP1‐overexpression (o/e), whereas RacGAP1‐siRNA inhibited cell malignancy. D,E, The same experiments were carried out on another serous EOC cell line, the Caov‐3 cell line, and showed consistent results. All experimental data were from three independent experiments and analyzed by Student's t test. Bar, mean ± standard deviation. * P < .05

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Migration, Western Blot, Negative Control, Over Expression, Standard Deviation

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Rac GTPase activating protein 1 (RacGAP1) positively regulate activation of RhoA and Erk proteins. A, Overexpression of RacGAP1 inhibited GTP‐Rac1 expression, whereas it enhanced GTP‐RhoA and p‐Erk levels. Namely, knockdown of RacGAP1 by siRNA increased GTP‐Rac1 level while it attenuated GTP‐RhoA and p‐Erk expression. B, Representative expression of RacGAP1 and p‐Erk in clinical epithelial ovarian cancer (EOC) tissue samples, which showed significant statistical correlations as revealed by χ 2 ‐test ( P = .003, [C])

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Activation Assay, Over Expression, Expressing, Knockdown

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Clinicopathological characteristics of EOC patients

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Expressing, Mutagenesis

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Prognostic parameters for OS in EOC patients by univariate and multivariate Cox proportional hazard model analyses

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Mutagenesis, Expressing

Journal: Cancer Science

Article Title: Rac GTPase activating protein 1 promotes oncogenic progression of epithelial ovarian cancer

doi: 10.1111/cas.13434

Figure Lengend Snippet: Prognostic parameters for PFS in EOC patients by univariate and multivariate Cox proportional hazard model analyses

Article Snippet: The plasmid constructs that express RacGAP1 (pcDNA‐RacGAP1), and the negative control vector plasmid (pcDNA) were purchased from GenScript (Nanjing, China). siRNA that targeted RacGAP1 and a scrambled negative control were purchased from GenePharma (Shanghai, China) with the following sequences: RacGAP1 siRNA #1: 5′‐GGU CUG ACU GAG ACA GGC CUG UAU A‐3′; RacGAP1 siRNA #2: 5′‐GCC AAG AAC UGA GAC AGA CAG UGU G‐3′; RacGAP1 siRNA #3: 5′‐CAA CUA AGC GAG GAG CAA A‐3′; Scrambled control siRNA: 5′‐UUC UCC GAA CGU GUA CGU‐3′.

Techniques: Mutagenesis, 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: Three different siRNAs, namely anti-RACGAP1 (si-RACGAP1-1/-2/-3) and control siRNA (si-Ctrl), were purchased from Ribobio (Guangzhou, Guangdong, 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: Three different siRNAs, namely anti-RACGAP1 (si-RACGAP1-1/-2/-3) and control siRNA (si-Ctrl), were purchased from Ribobio (Guangzhou, Guangdong, 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: Three different siRNAs, namely anti-RACGAP1 (si-RACGAP1-1/-2/-3) and control siRNA (si-Ctrl), were purchased from Ribobio (Guangzhou, Guangdong, China).

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

Journal: Molecular Oncology

Article Title: Long non‐coding RNA RACGAP1P promotes breast cancer invasion and metastasis via miR‐345‐5p/RACGAP1‐mediated mitochondrial fission

doi: 10.1002/1878-0261.12866

Figure Lengend Snippet: RACGAP1P was co‐expressed with RACGAP1 in breast cancer. (A) Co‐expression analysis of RACGAP1P and RACGAP1 in 460 microarray samples from TCGA dataset was visualized with cBioPortal. Pearson, R = 0.74. P < 0.01. (B)The mRNA expression level of RACGAP1 was revealed in breast cancer, performed on UALCAN online software. *** P < 0.001 (C and D) QRT‐PCR was performed to evaluate RACGAP1 mRNA expression in RACGAP1P overexpressing MDA‐MB‐231 and MCF7 cells. ** P < 0.01. (E and F) Western blot was carried on to evaluate RACGAP1 protein expression in RACGAP1P overexpressed MDA‐MB‐231 and MCF7 cells. ** P < 0.01.

Article Snippet: Proteins were transferred to poly(vinylidene difluoride) membranes (Millipore, Billerica, MA, USA) and immunoblotted with the indicated antibodies as follows: RACGAP1 (1 : 1000; AbSci, #AB32752, Nanjing, China), Drp1 (1 : 1000; CST, #5391, Danvers, MA, USA), p‐Drp1 (Ser161) (1 : 1000; CST, #4494), GAPDH (1 : 1000; Santa Cruz, # sc‐51097, Santa Cruz, CA, USA), Mfn1 (1 : 1000; Santa Cruz, # sc‐166644, ), Mfn2 (1 : 1000; CST, #9482).

Techniques: Expressing, Microarray, Software, Quantitative RT-PCR, Western Blot

Journal: Molecular Oncology

Article Title: Long non‐coding RNA RACGAP1P promotes breast cancer invasion and metastasis via miR‐345‐5p/RACGAP1‐mediated mitochondrial fission

doi: 10.1002/1878-0261.12866

Figure Lengend Snippet: RACGAP1P acted as a ceRNA titrating miR‐345‐5p and up‐regulating RACGAP1. (A) The correlation between RACGAP1P and miR‐345‐5p was evaluated by Pearson analysis in 25 breast cancer tissues. R = −0.4106. P = 0.0415. (B) The correlation between RACGAP1 and miR‐345‐5p was evaluated by Pearson analysis in 25 breast cancer tissues. R = −0.4538. P = 0.0227. (C and D) QRT‐PCR was used to detect RACGAP1P expression in MDA‐MB‐231 and MCF7 cells transfected with scramble sequence, miR‐345‐5p mimic, or miR345‐5p inhibitor. * P < 0.05. (E‐H) QRT‐PCR (E‐F) and Western blot (G‐H) were carried on to evaluate RACGAP1 expression in MDA‐MB‐231 and MCF7 cells transfected with scramble sequence, miR‐345‐5p mimic or miR345‐5p inhibitor. * P < 0.05. (I‐L) The expression level of RACGAP was measured by qRT‐PCR (I and J) and Western blot (K and L) in MDA‐MB‐231 and MCF7 cells transfected with indicated DNA or RNA. (M) Dual‐Luciferase reporter assay was performed in MDA‐MB‐231 and MCF7 cells co‐transfected with miR‐345‐5p mimics and pMIR‐REPORT carrying wild‐type or mutant target sequence at 3′‐UTR of the reporter gene. * P < 0.05. (N and O) RIP assay was conducted in MDA‐MB‐231 and MCF7 cells to examine miR‐345‐5p endogenously associated with RACGAP1P and RACGAP1. ** P < 0.01. Error bars, SD; Student’s t ‐test.

Article Snippet: Proteins were transferred to poly(vinylidene difluoride) membranes (Millipore, Billerica, MA, USA) and immunoblotted with the indicated antibodies as follows: RACGAP1 (1 : 1000; AbSci, #AB32752, Nanjing, China), Drp1 (1 : 1000; CST, #5391, Danvers, MA, USA), p‐Drp1 (Ser161) (1 : 1000; CST, #4494), GAPDH (1 : 1000; Santa Cruz, # sc‐51097, Santa Cruz, CA, USA), Mfn1 (1 : 1000; Santa Cruz, # sc‐166644, ), Mfn2 (1 : 1000; CST, #9482).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Sequencing, Western Blot, Luciferase, Reporter Assay, Mutagenesis

Journal: Molecular Oncology

Article Title: Long non‐coding RNA RACGAP1P promotes breast cancer invasion and metastasis via miR‐345‐5p/RACGAP1‐mediated mitochondrial fission

doi: 10.1002/1878-0261.12866

Figure Lengend Snippet: RACGAP1P‐induced cell invasion depended on RACGAP1. (A and B) Western blot analysis was used to detect the expression levels of RACGAP1, Drp1, and p‐Drp1 in MDA‐MB‐231 and MCF7 cells stably transfected with scramble sequence, RACGAP1 CDS, and RACGAP1 shRNA. * P < 0.05, ** P < 0.01. (C and D) Mitochondrial morphology staining (C) and transwell assays (D) were performed in MDA‐MB‐231 and MCF7 cells stably transfected with scramble sequence, RACGAP1, and RACGAP1 shRNA. Scale bar (white), 5 μm; scale bar (red), 50 μm. * P < 0.05, ** P < 0.01. (E and F) Western blot analysis was used to detect the expression levels of RACGAP1, Drp1, and p‐Drp1 in RACGAP1 knockdown MDA‐MB‐231 and MCF7 cells stably transfected with scramble sequence, RACGAP1P, and RACGAP1 sh‐re . ** P < 0.01. (G‐H) Mitochondrial morphology staining (G) and transwell assays (H) were performed in RACGAP1 knockdown MDA‐MB‐231 and MCF7 cells stably transfected with scramble sequence, RACGAP1P, and RACGAP1 sh‐re . Scale bar (white), 5 μm; scale bar (red), 50 μm. * P < 0.05, ** P < 0.01. Error bars, SD; Student’s t ‐test.

Article Snippet: Proteins were transferred to poly(vinylidene difluoride) membranes (Millipore, Billerica, MA, USA) and immunoblotted with the indicated antibodies as follows: RACGAP1 (1 : 1000; AbSci, #AB32752, Nanjing, China), Drp1 (1 : 1000; CST, #5391, Danvers, MA, USA), p‐Drp1 (Ser161) (1 : 1000; CST, #4494), GAPDH (1 : 1000; Santa Cruz, # sc‐51097, Santa Cruz, CA, USA), Mfn1 (1 : 1000; Santa Cruz, # sc‐166644, ), Mfn2 (1 : 1000; CST, #9482).

Techniques: Western Blot, Expressing, Stable Transfection, Transfection, Sequencing, shRNA, Staining

Journal: Molecular Oncology

Article Title: Long non‐coding RNA RACGAP1P promotes breast cancer invasion and metastasis via miR‐345‐5p/RACGAP1‐mediated mitochondrial fission

doi: 10.1002/1878-0261.12866

Figure Lengend Snippet: Potential mechanism of RACGAP1P implicated in breast cancer invasion and metastasis. In breast cancer cells, RACGAP1P could competitively bind to miR‐345‐5p that targeted RACGAP1 therefore up‐regulate RACGAP1. RACGAP1P overexpression promoted mitochondrial fission mediated cell invasion via RACGAP1P/miR‐345‐5p/RACGAP1/Drp1 network.

Article Snippet: Proteins were transferred to poly(vinylidene difluoride) membranes (Millipore, Billerica, MA, USA) and immunoblotted with the indicated antibodies as follows: RACGAP1 (1 : 1000; AbSci, #AB32752, Nanjing, China), Drp1 (1 : 1000; CST, #5391, Danvers, MA, USA), p‐Drp1 (Ser161) (1 : 1000; CST, #4494), GAPDH (1 : 1000; Santa Cruz, # sc‐51097, Santa Cruz, CA, USA), Mfn1 (1 : 1000; Santa Cruz, # sc‐166644, ), Mfn2 (1 : 1000; CST, #9482).

Techniques: Over Expression