Journal: Oncogene

Article Title: Upregulation of PIP3-Dependent Rac Exchanger 1 (P-Rex1) Promotes Prostate Cancer Metastasis

doi: 10.1038/onc.2009.30

Figure Lengend Snippet: P-Rex1 promotes prostate cancer cell migration and invasion via Rac activation. (a) CWR22Rv1 cells (left panel) or PC3-LN4 cells (right panel) were transfected with control GFP, GFP-tagged wild-type P-Rex1 or P-Rex1(-DH) mutant for 48 h, and then subjected to Transwell migration assay for 12 h (CWR22Rv1 cells) or 5 h (PC3-LN4) toward NIH3T3 CM. Transfected PC3-LN4 cells were selected by flow cytometry cell sorting based on GFP fluorescence prior to migration assays due to a low transfection efficiency (~25%). Bars show mean ± S.E. with *p<0.05 compared to control set at 100% (n=3). Inset: Expression of GFP-tagged P-Rex1 and P-Rex1(-DH) protein detected by western blot analysis using anti-GFP antibody. (b) CWR22Rv1 cells stably expressing wild-type P-Rex1, P-Rex1 “GEF-dead” mutant or control vector were subjected to migration and invasion assays for 12 h. Cells stably expressing wild-type P-Rex1 were pretreated without (none) or with 50 μM NSC for 15 min. Bars show mean ± S.E. with *p<0.01 compared to control (n=3). Inset: Western blot analysis of expression levels of P-Rex1 and its mutant. (c) P-Rex1 activates Rac1 in prostate cancer cells. The amount of activated GTP-bound Rac1 was measured by a Rac activation assay. The image shown is representative of 3 separate experiments. (d) P-Rex1 increases lamellipodia formation. Serum-starved CWR22Rv1 cells or PC-3 cells (positive control) were incubated with NIH3T3 CM for 5 h and then stained with rhodamine-conjugated phalloidin (red) for detection of F-actin.

Article Snippet: Additional reagents were NSC23766, Y27632 and anti-GFP antibody (EMD Biosciences); cell permeable C3 transferase and F-actin visualization kit (Cytoskeleton); Rac activation assay kit, Rho assay reagent, anti-Rac1 antibody (clone 23A8), and Immobilon-FL (Millipore); mouse anti-RhoA (26C4), and rabbit anti-HA and anti-β-actin antibodies (Santa Cruz); FBS (Hyclone); Lipofectamine TM 2000, AlexaF488-labeled secondary antibody, RPMI-1640 and DMEM (Invitrogen); EGF, Matrigel and fibronectin (BD Biosciences); CXCL12 (R&D Systems); and IRDye-labeled secondary antibodies (LI-COR Biosciences).

Techniques: Migration, Activation Assay, Transfection, Control, Mutagenesis, Transwell Migration Assay, Flow Cytometry, FACS, Fluorescence, Expressing, Western Blot, Stable Transfection, Plasmid Preparation, Positive Control, Incubation, Staining

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: Expression differences and diagnostic efficacy of key genes (A) Expression differences of key genes between tumor and normal samples (GEO: GSE60052 ). (B–E) ROC curves related to key genes (GEO: GSE60052 ). (F) Validation of expression differences of key genes (GEO: GSE149507 ). (G–J) ROC curves related to key genes (GEO: GSE149507 ). (K) UMAP plot of single-cell transcriptome data, illustrating the distribution of cell types in the SCLC microenvironment, including immune cells (T cells, B cells, macrophages, neutrophils, and DC) and SCLC subtypes (SCLC-A, SCLC-N, and SCLC-P), depicting their spatial arrangement. (L) UMAP plots showing the expression distribution of key genes ( GPX3 , RND2 , TRIM36 , and CAMK2N2 ) in SCLC single-cell transcriptomes (data source: Chan et al. ). Data are presented as mean ± SD. Statistical significance: ∗∗∗ p < 0.001. Statistical analyses were performed using R software. ROC curves and AUC values were calculated with the pROC package, and differences in AUC were compared using the DeLong test. ROC, receiver operating characteristic.

Article Snippet: RND2 , Proteintech , Cat#13844-1-AP; RRID: AB_2181817.

Techniques: Expressing, Diagnostic Assay, Biomarker Discovery, Single Cell, Software

Journal: iScience

Article Title: TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer

doi: 10.1016/j.isci.2026.115310

Figure Lengend Snippet: Survival analysis of patients based on key genes and their correlation with APM genes (A–D) Survival curves of groups with high and low expression of key genes ( GPX3 , CAMK2N2 , TRIM36 , and RND2 ) (George’s cohort ). (E) Scatterplot of the average expression of key genes vs. the average correlation with antigen presentation genes. (F and G) Scatterplots of the correlation between TRIM36, CAMK2N2 and HLA-A, HLA-B, HLA-C, B2M, respectively. Statistical significance was defined as p < 0.05. Statistical analyses were performed using R software. Survival analysis was conducted using the Kaplan-Meier method, and differences between groups were compared using the log rank test. Correlations were analyzed using Pearson’s correlation analysis. HLA, human leukocyte antigen; B2M, beta-2 microglobulin.

Article Snippet: RND2 , Proteintech , Cat#13844-1-AP; RRID: AB_2181817.

Techniques: Expressing, Immunopeptidomics, Software

Journal: Scientific reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells.

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Fig. 5. Rap1 overexpression reverses the expression of key proteins in CD147 knockdown mediated cell proliferation, apoptosis and EMT. (A, B) Western blot analysis of Rap1 and Rap1GAP protein levels in HCT116 and SW620 cells after CD147 knockdown (shCD147). β-actin served as a loading control. Data represent mean ± SD (n = 3; **P < 0.01, ***P < 0.001 vs. shNC, Student’s t-test). Original blots are presented in Supplementary Fig. 3. (C) qRT-PCR validation of Rap1 mRNA overexpression in CD147-knockdown cells. Expression normalized to β-actin (n = 3; ***P < 0.001 vs. HCT116 or SW620, Student’s t-test). (D, E) Western blot analysis of c-Myc, Bcl-2, Bax, N-cadherin, and E-cadherin protein levels in CD147-Knockdown cells with Rap1 overexpression (n = 3; *P < 0.05, **P < 0.01, ***P < 0.001 vs. shNC, #P < 0.05, ##P < 0.01, ###P < 0.001 vs. shCD147, one-way ANOVA). Original blots are presented in Supplementary Fig. 4.

Article Snippet: Supplier Antibody dilution used WB/IF Anti CD147 mouse monoclonal antibody 66443-1-AP ProteinTech Group, Inc. 1:5000 /1:200 Anti N-cadherin rabbit polyclonal antibody 22018-1-AP ProteinTech Group, Inc. 1:5000 Anti E-cadherin rabbit polyclonal antibody 20874-1-AP ProteinTech Group, Inc. 1:10000 Anti c-myc mouse monoclonal antibody 67,447-AP ProteinTech Group, Inc. 1:5000 Anti RAP1GAP rabbit polyclonal antibody 19174-1-AP ProteinTech Group, Inc. 1:2000 Anti RAP1 rabbit polyclonal antibody 14595-1-AP ProteinTech Group, Inc. 1:1000 Anti β-actin rabbit polyclonal antibody 20536-1-AP ProteinTech Group, Inc. 1:8000 HRP-conjugated Goat Anti-Mouse IgG(H + L) SA00001-1 ProteinTech Group, Inc. 1:10000 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) ab150116 Abcam 1:200 Table 1.

Techniques: Over Expression, Expressing, Knockdown, Western Blot, Control, Quantitative RT-PCR, Biomarker Discovery

Journal: Scientific reports

Article Title: CD147 regulates the Rap1 signaling pathway to promote proliferation, migration, and invasion, and inhibit apoptosis in colorectal cancer cells.

doi: 10.1038/s41598-025-98266-8

Figure Lengend Snippet: Fig. 7. Rap1 restores CD147-mediated migration and invasion in colorectal cancer cells. (A, C) Scratch wound healing assay in HCT116 cells. Healing rates were quantified at 24 h and 48 h (n = 3; ***P < 0.001 vs. shNC, ##P < 0.01, ###P < 0.001 vs. shCD147, one-way ANOVA). (B, D) Transwell migration and Matrigel invasion assays. Migrated/invaded cells were counted and normalized to shCtrl (n = 3; ***P < 0.001 vs. shNC, ###P < 0.001 vs. shCD147, one-way ANOVA). SW620 cells showed consistent trends. (E) Mechanistic diagram of CD147 promoting tumor progression through Rap1/Rap1GAP signaling in colorectal cancer cells.

Article Snippet: Supplier Antibody dilution used WB/IF Anti CD147 mouse monoclonal antibody 66443-1-AP ProteinTech Group, Inc. 1:5000 /1:200 Anti N-cadherin rabbit polyclonal antibody 22018-1-AP ProteinTech Group, Inc. 1:5000 Anti E-cadherin rabbit polyclonal antibody 20874-1-AP ProteinTech Group, Inc. 1:10000 Anti c-myc mouse monoclonal antibody 67,447-AP ProteinTech Group, Inc. 1:5000 Anti RAP1GAP rabbit polyclonal antibody 19174-1-AP ProteinTech Group, Inc. 1:2000 Anti RAP1 rabbit polyclonal antibody 14595-1-AP ProteinTech Group, Inc. 1:1000 Anti β-actin rabbit polyclonal antibody 20536-1-AP ProteinTech Group, Inc. 1:8000 HRP-conjugated Goat Anti-Mouse IgG(H + L) SA00001-1 ProteinTech Group, Inc. 1:10000 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) ab150116 Abcam 1:200 Table 1.

Techniques: Migration, Wound Healing Assay

Journal: PLoS ONE

Article Title: DNMT1 and HDAC2 Cooperate to Facilitate Aberrant Promoter Methylation in Inorganic Phosphate-Induced Endothelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0147816

Figure Lengend Snippet: The list of antibodies used for Western blotting.

Article Snippet: RASAL1 , 46–269 , 1:1000 , ProSci.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: DNMT1 and HDAC2 Cooperate to Facilitate Aberrant Promoter Methylation in Inorganic Phosphate-Induced Endothelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0147816

Figure Lengend Snippet: The sequences of primers used for real-time PCR.

Article Snippet: RASAL1 , 46–269 , 1:1000 , ProSci.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: DNMT1 and HDAC2 Cooperate to Facilitate Aberrant Promoter Methylation in Inorganic Phosphate-Induced Endothelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0147816

Figure Lengend Snippet: (A) Decreasing mRNA expression levels of RASAL1 upon the time of high Pi treatment after 24, 48 and 72 hours. Results were normalized to reference gene GAPDH (expression is presented as means ± s.d., n = 3 independent experiments, *P<0.05, **P<0.01, ***P<0.001). (B) Western blot confirming the decreased protein expression of RASAL1 in Pi treated cells. (C) MeDIP result showing the methylated promoter of RASAL1 along the treatment time, correlated with the reduced expression of RASAL1 in both mRNA and protein level. (D) Ras activity was measured by ELISA assay, untreated cells served as controls. Pi- treated cells showed the hyper-activation of total Ras.

Article Snippet: RASAL1 , 46–269 , 1:1000 , ProSci.

Techniques: Expressing, Western Blot, Methylated DNA Immunoprecipitation, Methylation, Activity Assay, Enzyme-linked Immunosorbent Assay, Activation Assay

Journal: PLoS ONE

Article Title: DNMT1 and HDAC2 Cooperate to Facilitate Aberrant Promoter Methylation in Inorganic Phosphate-Induced Endothelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0147816

Figure Lengend Snippet: Cells were treated with Pi 3mM for 72 hours. (A) MeDIP result showing the demethylated promoter of RASAL1 by using PFA, correlated with restored mRNA expression of RASAL1. (B) qPCR analysis showed the restored RASAL1 mRNA expression levels by using phosphate transporter inhibitor (PFA). Results were normalized to reference gene GAPDH (expression is presented as means ± s.d., n = 3 independent experiments, **P<0.01). (C) ELISA assay indicating the normalization of total Ras with combination of PFA or RAS inhibitor farnesylthiosalicylic acid (FTS). (D) bright-field images showing the morphology change of HCAEC cells cultured under normal control condition, high Pi conditions, and Pi combined with FTS or PFA. Scale bars 25 μm. (E) Western blot analysis showing the expression of endothelial cell marker CD31 and fibroblast cell marker S100A4 in HCAEC cells exposed to normal control condition, high Pi conditions, and Pi combined with FTS or PFA (F) qRT-PCR data showing the mRNA expression levels of EndMT transcriptional factors (SNAIL, SLUG, and TWIST) and FSP1 in HCAEC cells under four conditions indicated above. Results were normalized to reference gene GAPDH (expression is presented as means ± s.d., n = 3 independent experiments, ***P<0.001).

Article Snippet: RASAL1 , 46–269 , 1:1000 , ProSci.

Techniques: Methylated DNA Immunoprecipitation, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control, Western Blot, Marker, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: DNMT1 and HDAC2 Cooperate to Facilitate Aberrant Promoter Methylation in Inorganic Phosphate-Induced Endothelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0147816

Figure Lengend Snippet: (A) Simplified schematic showing the human RASAL1 promoter along with exons (black boxes), translational start site (black arrow), locations of RASAL1 chip primer for amplicon1 and amplicon2, and amplicon2 primers serve as negative control chip primer. (B-C) The binding properties of HDAC2 and DNMT1 to the RASAL1 promoter region were analyzed by chromatin immunoprecipitation (ChIP) assay and detected by qRT-PCR in Pi treated (B) or in control cells (C). IgG purified from the same species serve as negative control for ChIP (expression are presented as means ± s.d., n = 3 independent experiments, **P<0.01, ***P<0.001, n.s. no significance).

Article Snippet: RASAL1 , 46–269 , 1:1000 , ProSci.

Techniques: Negative Control, Binding Assay, Chromatin Immunoprecipitation, Quantitative RT-PCR, Control, Purification, Expressing

Journal: PLoS ONE

Article Title: DNMT1 and HDAC2 Cooperate to Facilitate Aberrant Promoter Methylation in Inorganic Phosphate-Induced Endothelial-Mesenchymal Transition

doi: 10.1371/journal.pone.0147816

Figure Lengend Snippet: In the physiological conditions, the CpG islands located in RASAL1 promoter are unmethylated (top panel) as indicated by open circles, and RASAL1 is transcriptional active. Under pathological conditions, initially when endothelial cells are exposed to stimulus, such as TGFβ1 or high concentration of Pi resulting in RASAL1 silencing through condensed chromatin structure at RASAL1 promoter mediated by HDAC2. While the RASAL1 promoter remains unmethylated (middle panel) and RASAL1 is transiently silenced. When the cells are continuously exposed to the stimulus, the CpG islands located in RASAL1 promoter are methylated (indicated by filled circles) by DNMT1 recruited through the interaction with HDAC2. Therefore, RASAL1 is permanently silenced due to promoter hypermethylation (lower panel).

Article Snippet: RASAL1 , 46–269 , 1:1000 , ProSci.

Techniques: Concentration Assay, Methylation