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: Pflugers Archiv

Article Title: The RyR2-P2328S mutation downregulates Na v 1.5 producing arrhythmic substrate in murine ventricles

doi: 10.1007/s00424-015-1750-0

Figure Lengend Snippet: MAP traces obtained from the epicardium of the left ventricle in WT and RyR2 S/S hearts during S1S2 pacing highlighting typical traces of either normal activity or arrhythmogenesis. All WT hearts entered the refractory period without displaying any episodes of arrhythmia, as defined by two or more non-stimulated APs ( a ); however, one heart displayed the occurrence of a singular ectopic (one non-stimulated AP) ( b ). Multiple arrhythmic events were observed in RyR2 S/S hearts including short non-sustained ventricular tachycardia (NSVT) ( c ), polymorphic tachycardia following a previously imposed S2 extrastimulus ( d ), monomorphic ventricular tachycardia (VT) ( e ) and episodes of VT which deteriorated into ventricular fibrillation (VF) (F). The small black circles indicate the timing of stimuli

Article Snippet: Hearts were paced at twice their excitation threshold voltage using a bipolar platinum-coated stimulating electrode placed on the ventricular septum connected to a DS2A-Mk.II stimulator (Digitimer).

Techniques: Activity Assay

Journal: Pflugers Archiv

Article Title: The RyR2-P2328S mutation downregulates Na v 1.5 producing arrhythmic substrate in murine ventricles

doi: 10.1007/s00424-015-1750-0

Figure Lengend Snippet: Typical MAP recordings obtained from the left ventricular epicardium of WT and RyR2 S/S during dynamic pacing. Traces from WT ( left ) and RyR2 S/S at progressively decreasing BCLs: 124 ( a ), 99 ( b ), 84 ( c ), 74 ( d ) and 54 ms ( e ). If a heart entered 2:1 block, the protocol was terminated (E). Traces are displayed along a common horizontal timescale. The vertical scale was normalized to a standard AP deflection at a BCL of 134 ms. Small black circles above each trace indicate the timing of stimuli

Article Snippet: Hearts were paced at twice their excitation threshold voltage using a bipolar platinum-coated stimulating electrode placed on the ventricular septum connected to a DS2A-Mk.II stimulator (Digitimer).

Techniques: Blocking Assay

Journal: Pflugers Archiv

Article Title: The RyR2-P2328S mutation downregulates Na v 1.5 producing arrhythmic substrate in murine ventricles

doi: 10.1007/s00424-015-1750-0

Figure Lengend Snippet: Total Cx43 expression in WT and RyR2 S/S ventricles. a Representative blots of Cx43 and GAPDH expression in WT and RyR2 S/S ventricular tissue and b the mean (± SEM) Cx43 expression normalized to GAPDH expression. Cx43 expression was similar between WT and RyR2 S/S (0.59 ± 0.07 and 0.79 ± 0.1, respectively; P = 0.167, n = 4) ventricles

Article Snippet: Hearts were paced at twice their excitation threshold voltage using a bipolar platinum-coated stimulating electrode placed on the ventricular septum connected to a DS2A-Mk.II stimulator (Digitimer).

Techniques: Expressing

Journal: Pflugers Archiv

Article Title: The RyR2-P2328S mutation downregulates Na v 1.5 producing arrhythmic substrate in murine ventricles

doi: 10.1007/s00424-015-1750-0

Figure Lengend Snippet: Western blots of Na v 1.5 expression in whole tissue and membrane fraction samples from WT and RyR2 S/S ventricles. Ventricular Na v 1.5 expression was decreased in RyR2 S/S compared to WT, both in the whole tissue (1.17 ± 0.20; n = 6, vs 1.69 ± 0.15 n = 7, respectively, P = 0.048) and in the membrane fraction (0.91 ± 0.13; n = 4, vs 2.06 ± 0.33; n = 4, respectively, P = 0.006). This suggested a greater proportional reduction in membrane relative to total Na v 1.5 expression in RyR2 S/S . Symbols denote significant differences between genotypes * P < 0.05, ** P < 0.01

Article Snippet: Hearts were paced at twice their excitation threshold voltage using a bipolar platinum-coated stimulating electrode placed on the ventricular septum connected to a DS2A-Mk.II stimulator (Digitimer).

Techniques: Western Blot, Expressing, Membrane

Journal: Pflugers Archiv

Article Title: The RyR2-P2328S mutation downregulates Na v 1.5 producing arrhythmic substrate in murine ventricles

doi: 10.1007/s00424-015-1750-0

Figure Lengend Snippet: Loose patch clamp recordings of I Na activation in WT and RyR2 S/S ventricles. a Representative currents in response to depolarizing steps increased from 20 to 120 mV in voltage-clamped WT and RyR2 S/S ventricular tissue. b Peak inward current (mean ± SEM) elicited at each voltage step for WT ( n = 6) and RyR2 S/S ( n = 12) ventricles. c The maximum current recorded during each complete voltage step protocol (mean ± SEM) was larger in the WT than the RyR2 S/S ventricles, P < 0.0047. The asterisks denote significant differences between genotypes of P < 0.01

Article Snippet: Hearts were paced at twice their excitation threshold voltage using a bipolar platinum-coated stimulating electrode placed on the ventricular septum connected to a DS2A-Mk.II stimulator (Digitimer).

Techniques: Patch Clamp, Activation Assay

Journal: Molecules

Article Title: Vincamine Ameliorates Epithelial-Mesenchymal Transition in Bleomycin-Induced Pulmonary Fibrosis in Rats; Targeting TGF-β/MAPK/Snai1 Pathway

doi: 10.3390/molecules28124665

Figure Lengend Snippet: Effect of vincamine on fibronectin ( A ), N-cadherin ( B ), and collagen ( C ) levels in lung tissues. Bars represent mean ± SD. Significant difference was analyzed by one-way ANOVA test followed by post hoc Dunnett test, where * p < 0.001, compared to sham group, and # p < 0.001, compared to BLM-induced group.

Article Snippet: In lung tissue homogenates, levels of fibronectin, N-cadherin, and collagen were also assessed, according to the manufacturer’s instructions, utilizing rat fibronectin ELISA kit (#MBS761397, MyBioSource, CA, USA), N-cadherin ELISA kit (#KOA0665, Rockland immunochemical Inc., Royersford, PA, USA), and collagen type I ELISA kit (#MBS262647, MyBioSource, CA, USA), respectively.

Techniques: