Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: FAM65A binds to Ras and activates the Ras/ERK signaling to mediate RSK activation (A) The volcano plot analysis results for the FAM65A high-expression and low-expression groups from the TCGA database were shown. (B) The KEGG and GO results were shown. (C) The GSEA results were shown. (D) GSEA on DEGs between the FAM65A high-expression group and low-expression group in the Reactome database were shown. (E) IP was performed to detect the binding of FAM65A and Ras/p-RSK. (F) IP was performed to detect the binding of Ras and FAM65A/p-RSK. (G) Immunofluorescence was performed to detect the co-localization of FAM65A and Ras. Scale bars, 20 μm. (H) Western blot analysis the Ras and p -ERK expression in FAM65A knockdown or overexpression cells. Data are presented as mean ± SEM of biologically independent experiments.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Activation Assay, Expressing, Binding Assay, Immunofluorescence, Western Blot, Knockdown, Over Expression

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: Ras/ERK signaling activation was indispensable for FAM65A-mediated RSK activation and CRC progression (A) Western blot analysis of Ras and p -ERK expression in HCT116-FAM65A cells treated with 10 μM Abd-7, or without treatment. (B) Results from the CCK8 cell proliferation assay conducted on HCT116-FAM65A cells with and without the application of Abd-7, n = 3, ∗∗∗ p < 0.001. (C) Colony formation assay performed on HCT116-FAM65A cells treated with Abd-7 or not. (D) Quantitative analysis of the colony formation assay results, n = 3, ∗∗∗ p < 0.001. (E) Results from the EdU assay conducted on HCT116-FAM65A cells with and without the application of Abd-7. Scale bars, 100 μm. (F) Quantitative analysis of the EdU assay results, n = 3, ∗∗∗ p < 0.001. (G) Western blot analysis of Ki-67, cleaved Caspase 3, Bcl-2, and Bax expression in HCT116-FAM65A cells treated with Abd-7 or not. (H) Results from the apoptosis assay conducted on HCT116-FAM65A cells treated with Abd-7 or not. Scale bars, 50 μm. (I) Quantitative analysis of the apoptosis experiments, n = 3, ∗∗∗ p < 0.001. (J) Results from the Transwell migration assay conducted on HCT116-FAM65A cells with and without the application of Abd-7. Scale bars, 50 μm. (K) Quantitative analysis of the Transwell migration assay results, n = 3, ∗∗∗ p < 0.001. (L) Results from the wound healing assay performed on HCT116-FAM65A cells treated with Abd-7 or not. Scale bars, 50 μm. (M) Quantitative analysis of the wound healing assay results, n = 3, ∗∗∗ p < 0.001. (N) Western blot analysis the expression of EMT markers in HCT116-FAM65A cells treated with Abd-7 or not. (O) Proposed model of FAM65A in CRC progression. Data are presented as mean ± SEM of biologically independent experiments.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Activation Assay, Western Blot, Expressing, Proliferation Assay, Colony Assay, EdU Assay, Apoptosis Assay, Transwell Migration Assay, Wound Healing Assay

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: Knockdown of FAM65A inhibits tumor progression in vivo (A) LOVO-shCtrl and LOVO-shFAM65A cells were administered into the fourth fat pad of nude mice, and the resulting tumor growth curves were subsequently generated, n = 5, ∗ p < 0.05. (B) The tumors excised from mice across various experimental groups are presented. (C) Hematoxylin and Eosin (HE) staining results of lung tissue from the different groups is displayed. (D) A quantitative analysis of metastatic lung nodules is provided, n = 5, ∗∗ p < 0.01. (E) IHC results for FAM65A, Ki-67, p -RSK, p -ERK, Ras, N-cadherin, vimentin, cleaved Caspase 3, ZO-1, and E-cadherin in tumor tissues are illustrated. (F) A quantitative analysis of the IHC results is included. Data are presented as mean ± SEM of biologically independent experiments, n = 5, ∗∗∗ p < 0.001.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Knockdown, In Vivo, Generated, Staining

Journal: Journal of Inflammation Research

Article Title: CCL20 Expression via AKT-ERK1/2-AP1 Pathway in Mycoplasma Pneumoniae Infection: Implications for EMT and Cell Migration

doi: 10.2147/JIR.S512408

Figure Lengend Snippet: The ERK1/2 signaling pathway is involved in regulating the expression of CCL20. THP-1 cells were infected with M. pneumoniae at MOIs of 0, 10, 50, or 100 for 1 h, and protein expression levels were assessed by Western blot ( A and B ). In inhibitor experiments, cells were pretreated with 30 µM ERK1/2 inhibitor PD98059 for 1 h before infection with M. pneumoniae . The inhibitory effect was detected by Western blot ( C and D ). The levels of CCL20 mRNA and protein were measured by RT-qPCR ( E ) and ELISA ( F ), respectively. Additionally, cells were pretreated with 50 µM LY294002 prior to infection, and the inhibitory effect on ERK1/2 was detected by Western blot ( G and H ). Representative results from three independent experiments are shown. * p <0.05, ** p <0.01, **** p <0.0001. * indicates significance compared to the control group.

Article Snippet: Subsequently, the membrane was incubated overnight at 4°C with primary antibodies, including HY-P80276 P-AKT, HY-P80009 AKT, HY-P80813 P-ERK1/2, HY-P80393 ERK1/2, HY-P80084 c-jun, HY-P80616 c-fos, and HY-P80112 E-cadherin (all from MCE, USA, diluted at 1:800), as well as HY-P80438 β-actin (from MCE, USA, diluted at 1:10,000) and D155001 p-c-jun, D151325 p-c-fos, and D121239 snail (from Sangon Biotech, China, diluted at 1:800).

Techniques: Expressing, Infection, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Dental Sciences

Article Title: Overexpression of sprouty 1 protein in human oral squamous cell carcinogenesis

doi: 10.1016/j.jds.2020.07.013

Figure Lengend Snippet: Western blot analyses: phosphor-ERK and total-ERK in human oral cancer cell lines as compared with a primary culture of normal oral mucosa (HOK). Upregulation of phosphor-ERK (normalized to total-ERK) in OECM1 and Ca922 cell lines as compared with HOK, and a slightly decreased expression in SAS as compared with HOK. Results were quantified using densitometric analysis, normalized to the level of β-actin, and expressed as a fold change relative to the normal oral mucosa. Bars represent means ± standard deviation of the mean (∗ P < 0.05). A representative result of three independent experiments is shown.

Article Snippet: Further, samples were analyzed using 10% SDS-PAGE (Sigma-Aldrich) gels, and the proteins were transmitted onto a PVDF membrane (Sigma-Aldrich) using Bio-Rad's transblot with primary antibodies against phosphor-ERK (Boster Biological Technology, CA, USA; Cat. No. P00104; 1:1000) and total-ERK (Boster Biological Technology; Cat. No. P00104; 1:1000), with species specificity for human tissues and an observed molecular weight of 42–44 kDa; and β-actin (Sigma-Aldrich; 1:1000), followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (Sigma-Aldrich; 1:5000).

Techniques: Western Blot, Expressing, Standard Deviation