Journal: iScience

Article Title: DAB2IP suppresses invadopodia formation through destabilizing ALK by interacting with USP10 in breast cancer

doi: 10.1016/j.isci.2023.107606

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Article Snippet: Rabbit anti-p-Src family(Tyr416) , Cell signaling technology , Cat# 59548, RRID: AB_2936373.

Techniques: Virus, Recombinant, Magnetic Beads, CCK-8 Assay, Ab Array, Mutagenesis, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Expressing, CRISPR, Knock-Out, Software

Journal: Nature Communications

Article Title: Fibroblast growth factor 18 alleviates stress-induced pathological cardiac hypertrophy in male mice

doi: 10.1038/s41467-023-36895-1

Figure Lengend Snippet: a The corresponding Src family kinases correlated with the non-hypertrophic myocardium and hypertrophic myocardium were visualized by the heatmap in the normal and ISO treatment samples in GSE18801 dataset. The darker shade of red or blue represents the higher correlation level. b Circle plot depicting the important signal pathways associated with related genes. c NRCMs were treated with ISO (10 μM) in the presence or absence of FGF18 (50 ng/ml) for 48 h. Protein bands at ~60 kDa were excised from the SDS-PAGE gels for LC-MS/MS analysis. d NRCMs were treated with FGF18 (50 ng/ml) for 10 min. The cell lysates were subjected to immunoprecipitation with FGFR3 antibody, followed by immunoblotting with the FYN antibody. Cell lysates were also subjected to immunoprecipitation with IgG as negative control. n = 4. e NRCMs were treatment with FGF18 (50 ng/ml) for 10 min. The cell lysates were subjected to immunoprecipitation with FYN antibody, followed by immunoblotting with the p-SrcY416 and FGFR3 antibodies. Cell lysates were also subjected to immunoprecipitation with IgG as a negative control. n = 4. All numbers ( n ) are biologically independent experiments. Source data are provided as a Source data file.

Article Snippet: The primary antibodies were p-p38 (CST, 4511, 1:1000), p38 (CST, 8690, 1:2000), p-Erk (CST, 9101, 1:2000), Erk (CST, 9102, 1:2000), p-JNK (CST, 4668, 1:1000), JNK (Abcam, ab179461, 1:2000), Bax (CST, 2772, 1:2000), Bcl-2 (Santa cruz, sc-7382, 1:1000), FGF1 (Abcam, ab207321, 1:1000), FGF2 (Santa cruz, sc-74412, 1:1000), FGF3 (Santa cruz, sc-135, 1:1000), FGF5 (Santa cruz, sc-376264, 1:1000), FGF9 (Santa cruz, sc-373716, 1:1000), FGF13 (Affinity biosciences, DF4699, 1:1000), FGF16 (Santa cruz, sc-390547, 1:1000), FGF18 (Santa cruz, sc-393471, 1:1000), FGFR3 (Santa cruz, sc-390423, 1:1000), FYN (Santa cruz, sc-365913, 1:1000), p-Src (CST, 2101, 1:1000), 3-NT (Abcam, ab61392, 1:1000), NOX4 (Origene, TA349083, 1:1000), p22 phox (Santa cruz, sc-271968, 1:1000).

Techniques: SDS Page, Liquid Chromatography with Mass Spectroscopy, Immunoprecipitation, Western Blot, Negative Control

Journal: Nature Communications

Article Title: Fibroblast growth factor 18 alleviates stress-induced pathological cardiac hypertrophy in male mice

doi: 10.1038/s41467-023-36895-1

Figure Lengend Snippet: a , b The mice were injected with AAV9-LacZ or AAV9-cTnT-FGF18 intravenously before Sham or TAC operation. a Western blotting with p-FYN (IP: FYN, IB: p-Src) and FYN antibody. Quantification of relative FYN activity and protein expression levels (right panel). n = 5. Data represent means ± SD. one-way ANOVA with Tukey multiple comparisons test. b qRT-PCR analysis of FYN levels. n = 5. Data represent means ± SD. one-way ANOVA with Tukey multiple comparisons test: n.s. = not significant. c – i AAV9-cTnT-FGF18 and control vector were injected intravenously into tail veins of 6 weeks old male C57BL/6J mice, respectively. One week after the injection, these mice were intravenously injected with the adeno-associated virus (AAV9-cTnT-sh-FYN; AAV9-Scramble) into tail veins. One week after the injection, these mice were subjected to Sham or TAC operation for 6 weeks. c Histological analyses of the HE staining and WGA staining ( n = 5. Scale bar = 0.6 mm for upper HE staining; scale bar = 20 μm for lower WGA staining). Statistical results for the sectional cell area (right panel). n = 5. d Real-time quantitative PCR assays. n = 5. e Statistical results for the ratios of HW/BW in the indicated groups. n = 5. f Echocardiographic data for LVEF and LVFS are shown. n = 5. g PSR staining and Quantification (right panel). n = 5. Scale bar = 450 μm, and then zoom in 5 times. h Left ventricular collagen quantification by hydroxyproline assay (μg/mg). n = 5. i Levels of the oxidative damage marker 3-NT. The quantitative analysis of protein immunoblots (right panel), n = 5. j Total ROS levels (by DCFH-DA probe) were quantified. n = 4. k Hydrogen peroxide levels (by Amplex Red assay) quantified in different groups. n = 5. One-way ANOVA was followed by a post hoc Fisher’s comparison test. l Quantification of relative protein levels. n = 5. c – i All quantitative data are reported as means ± SD, one-way ANOVA with Tukey multiple comparisons test. All numbers ( n ) are biologically independent animals. Source data are provided as a Source data file.

Article Snippet: The primary antibodies were p-p38 (CST, 4511, 1:1000), p38 (CST, 8690, 1:2000), p-Erk (CST, 9101, 1:2000), Erk (CST, 9102, 1:2000), p-JNK (CST, 4668, 1:1000), JNK (Abcam, ab179461, 1:2000), Bax (CST, 2772, 1:2000), Bcl-2 (Santa cruz, sc-7382, 1:1000), FGF1 (Abcam, ab207321, 1:1000), FGF2 (Santa cruz, sc-74412, 1:1000), FGF3 (Santa cruz, sc-135, 1:1000), FGF5 (Santa cruz, sc-376264, 1:1000), FGF9 (Santa cruz, sc-373716, 1:1000), FGF13 (Affinity biosciences, DF4699, 1:1000), FGF16 (Santa cruz, sc-390547, 1:1000), FGF18 (Santa cruz, sc-393471, 1:1000), FGFR3 (Santa cruz, sc-390423, 1:1000), FYN (Santa cruz, sc-365913, 1:1000), p-Src (CST, 2101, 1:1000), 3-NT (Abcam, ab61392, 1:1000), NOX4 (Origene, TA349083, 1:1000), p22 phox (Santa cruz, sc-271968, 1:1000).

Techniques: Injection, Western Blot, Activity Assay, Expressing, Quantitative RT-PCR, Plasmid Preparation, Staining, Real-time Polymerase Chain Reaction, Hydroxyproline Assay, Marker, Amplex Red Assay

Journal: Pharmaceutics

Article Title: Licochalcone A Suppresses Renal Cancer Cell Proliferation and Metastasis by Engagement of Sp1-Mediated LC3 Expression

doi: 10.3390/pharmaceutics15020684

Figure Lengend Snippet: LicA inhibits metastasis via the FAK/Src signaling pathway of RCC cells. Different LicA concentrations (0, 6.25, 12.5, and 12.5 µM) were applied to ACHN cells, and the cells were then left to react for 24 h. ( A ) Western blotting was utilized to examine the expression of p-FAK, t-FAK, p-Src, and t-Src in cell lysates. β-actin served as a internal control. The ACHN cells were treated with 12.5 µM LicA in the presence or absence of ( B ) a FAK inhibitor (PF) or ( C ) an Src inhibitor (PP2) for 24 h. The cell lysates were examined using Western blotting to measure the protein levels of p-FAK, t-FAK, and LC3B. ( D , E ) Next, migration and Matrigel-invasion assays were used to assess migration and invasion. Data represent the mean ± SD of at least three separate experiments. ** p < 0.01, compared to un-treated control (0 µM); # p < 0.05, compared with LicA-treated alone.

Article Snippet: Anti-phosphoryl-FAK (p-FAK), total-FAK (t-FAK), phosphoryl-Src (p-Src) and total-Src (t-Src) antibodies were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Western Blot, Expressing, Migration

Journal: Pharmaceutics

Article Title: Licochalcone A Suppresses Renal Cancer Cell Proliferation and Metastasis by Engagement of Sp1-Mediated LC3 Expression

doi: 10.3390/pharmaceutics15020684

Figure Lengend Snippet: LicA inhibits RCC tumorigenesis and metastasis via the downregulation of the phosphoryl-Src/-FAK pathway. It does so by decreasing the Sp1 transcription activity, resulting in suppressed LC3B expression.

Article Snippet: Anti-phosphoryl-FAK (p-FAK), total-FAK (t-FAK), phosphoryl-Src (p-Src) and total-Src (t-Src) antibodies were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Activity Assay, Expressing