Journal: Biomedicines

Article Title: CARD14 Signalling Ensures Cell Survival and Cancer Associated Gene Expression in Prostate Cancer Cells.

doi: 10.3390/biomedicines10082008

Figure Lengend Snippet: Figure 1. High CARD14 expression correlates with aggressive cancer in human PCa patients. (A) Kaplan–Meier curve of the probability of disease-free survival for PCa patients with low or high CARD14, CARD10, MALT1, or BCL10 RNA level, based on the TCGA RNAseq dataset. p value generated by the Log-rank test is indicated. (B,C) CARD14 expression in normal and tumour (paired (C)) PCa tissue, based on TCGA data. (D–F) CARD14 expression as a function of lymph node metastasis (D) (no: no presence of metastasis in lymph nodes; micro: presence of micrometastasis in lymph nodes); tumour metastasis (E) (P: primary tumour, M: metastatic tumour) or biochemical recurrence (F) for PCa, based on TCGA data (D,F) or MSKCC data (E). p value generated by the Welsh t-test is indicated as * p < 0.05, *** p < 0.001. (G) CARD14 mRNA expression as a function of proliferation score.

Article Snippet: The following antibodies were used: anti-MALT1 polyclonal antibody (ab33921, Abcam, Amsterdam, Netherlands), anti-CARD14 polyclonal antibody (10400-1-AP, ProteinTech, Manchester, United Kingdom), anti-BCL10 monoclonal antibody (sc-5273, Santa-Cruz, Heidelberg, Germany), anti-c-Jun monoclonal antibody (sc-74543, Santa Cruz, Heidelberg, Germany), anti-phospho-c-Jun (Ser63) polyclonal antibody (9261, Cell Signaling Technology, MA, USA), anti-IκBα polyclonal antibody (sc-371, Santa Cruz, Heidelberg, Germany), anti-phospho-IκBα (Ser32/36) monoclonal antibody (9246, Cell Signaling Technology, MA, USA), anti-Akt polyclonal antibody (9272, Cell Signalling, MA, USA), antiphospho-Akt (Ser473) polyclonal antibody (9271, Cell Signalling, MA, USA), anti-CYLD monoclonal antibody (sc-74435, Santa Cruz, Heidelberg, Germany), anti-A20 monoclonal antibody (sc-166692, Santa Cruz, Heidelberg, Germany), and anti-actin monoclonal antibody (MP6472J, MP Biomedicals, Eschwege, Germany).

Techniques: Expressing, Generated

Journal: Biomedicines

Article Title: CARD14 Signalling Ensures Cell Survival and Cancer Associated Gene Expression in Prostate Cancer Cells.

doi: 10.3390/biomedicines10082008

Figure Lengend Snippet: Figure 2. CARD14 is essential for PCa cell survival. (A) CARD10 and CARD14 expression profile in PCa cell lines analysed by western blot. Actin expression was used as a loading control. (B) Cell death in function of time upon knock-down of CARD14 in LNCaP cells. The number of dead cells was quantified using SYTOX green staining and the Incucyte Live-Cell Analysis System. Results are representative of three independent experiments. Error bars represent means ± SEs.

Article Snippet: The following antibodies were used: anti-MALT1 polyclonal antibody (ab33921, Abcam, Amsterdam, Netherlands), anti-CARD14 polyclonal antibody (10400-1-AP, ProteinTech, Manchester, United Kingdom), anti-BCL10 monoclonal antibody (sc-5273, Santa-Cruz, Heidelberg, Germany), anti-c-Jun monoclonal antibody (sc-74543, Santa Cruz, Heidelberg, Germany), anti-phospho-c-Jun (Ser63) polyclonal antibody (9261, Cell Signaling Technology, MA, USA), anti-IκBα polyclonal antibody (sc-371, Santa Cruz, Heidelberg, Germany), anti-phospho-IκBα (Ser32/36) monoclonal antibody (9246, Cell Signaling Technology, MA, USA), anti-Akt polyclonal antibody (9272, Cell Signalling, MA, USA), antiphospho-Akt (Ser473) polyclonal antibody (9271, Cell Signalling, MA, USA), anti-CYLD monoclonal antibody (sc-74435, Santa Cruz, Heidelberg, Germany), anti-A20 monoclonal antibody (sc-166692, Santa Cruz, Heidelberg, Germany), and anti-actin monoclonal antibody (MP6472J, MP Biomedicals, Eschwege, Germany).

Techniques: Expressing, Western Blot, Control, Knockdown, Staining, Cell Analysis

Journal: Biomedicines

Article Title: CARD14 Signalling Ensures Cell Survival and Cancer Associated Gene Expression in Prostate Cancer Cells.

doi: 10.3390/biomedicines10082008

Figure Lengend Snippet: Figure 3. CARD14 expression correlates with DNA repair gene enrichment. (A) Gene ontology analysis for enrichment of biological processes on all genes whose expression showed a significant correlation with CARD14 expression based on gene expression (RSEM) in TCGA prostate adenocarci- noma (PRAD) patients. (B) Gene set enrichment analysis for enrichment of the Hallmark DNA repair gene set in genes ranked by positive correlation with CARD14 expression in TCGA PRAD patients (NES: Normalised Enrichment Score, FDR: False Discovery Rate). (C) Pearson correlation analysis between DNA damage response (DDR) pathway activity and CARD14 expression in TCGA PRAD patients. (D–G) CARD14 mRNA expression as a function of disease progression after radiotherapy (D), fraction genome altered (E), microsatellite instability (F) and aneuploidy score (G) in PCa patients, based on TCGA data. p value in (D) was generated by Welch t-test, * p < 0.05.

Article Snippet: The following antibodies were used: anti-MALT1 polyclonal antibody (ab33921, Abcam, Amsterdam, Netherlands), anti-CARD14 polyclonal antibody (10400-1-AP, ProteinTech, Manchester, United Kingdom), anti-BCL10 monoclonal antibody (sc-5273, Santa-Cruz, Heidelberg, Germany), anti-c-Jun monoclonal antibody (sc-74543, Santa Cruz, Heidelberg, Germany), anti-phospho-c-Jun (Ser63) polyclonal antibody (9261, Cell Signaling Technology, MA, USA), anti-IκBα polyclonal antibody (sc-371, Santa Cruz, Heidelberg, Germany), anti-phospho-IκBα (Ser32/36) monoclonal antibody (9246, Cell Signaling Technology, MA, USA), anti-Akt polyclonal antibody (9272, Cell Signalling, MA, USA), antiphospho-Akt (Ser473) polyclonal antibody (9271, Cell Signalling, MA, USA), anti-CYLD monoclonal antibody (sc-74435, Santa Cruz, Heidelberg, Germany), anti-A20 monoclonal antibody (sc-166692, Santa Cruz, Heidelberg, Germany), and anti-actin monoclonal antibody (MP6472J, MP Biomedicals, Eschwege, Germany).

Techniques: Expressing, Gene Expression, Activity Assay, Biomarker Discovery, Generated

Journal: Biomedicines

Article Title: CARD14 Signalling Ensures Cell Survival and Cancer Associated Gene Expression in Prostate Cancer Cells.

doi: 10.3390/biomedicines10082008

Figure Lengend Snippet: Figure 4. MALT1 protease activity is constitutively active in PCa cells, but PCa cell survival only requires MALT1 scaffold activity and not its catalytic activity. (A) Schematic representation of CARD14 signalling in epithelial cells. Activation of CARD14 by a yet to be defined upstream stimulus or a gain-of-function mutation triggers formation of a CARD14–BCL10–MALT1 complex leading to NF-κB- and MAPK-mediated gene expression. Additionally, CBM formation unlocks the ability of MALT1 to cleave several substrates that further regulate downstream events. (B,C) Cell death in function of time upon knock-down of MALT1 or BCL10 in LNCaP cells (B), PC3 and DU145 cells (C), as indicated. The number of dead cells was quantified using SYTOX green staining and the Incucyte Live-Cell Analysis System. (D) The indicated PCa cell lines were left untreated (−) or treated (+) with MALT1 inhibitor (1 µM) for 48 h. CYLD and A20 cleavage (indicated with an arrow head), as well as MALT1 expression was visualised by western blot of the corresponding cell extracts. (E) Cell death in function of time upon MALT1 inhibition in LNCaP cells, analysed as in (B). Results are representative of three independent experiments. Error bars (B,C,E) represent means ± SEs.

Article Snippet: The following antibodies were used: anti-MALT1 polyclonal antibody (ab33921, Abcam, Amsterdam, Netherlands), anti-CARD14 polyclonal antibody (10400-1-AP, ProteinTech, Manchester, United Kingdom), anti-BCL10 monoclonal antibody (sc-5273, Santa-Cruz, Heidelberg, Germany), anti-c-Jun monoclonal antibody (sc-74543, Santa Cruz, Heidelberg, Germany), anti-phospho-c-Jun (Ser63) polyclonal antibody (9261, Cell Signaling Technology, MA, USA), anti-IκBα polyclonal antibody (sc-371, Santa Cruz, Heidelberg, Germany), anti-phospho-IκBα (Ser32/36) monoclonal antibody (9246, Cell Signaling Technology, MA, USA), anti-Akt polyclonal antibody (9272, Cell Signalling, MA, USA), antiphospho-Akt (Ser473) polyclonal antibody (9271, Cell Signalling, MA, USA), anti-CYLD monoclonal antibody (sc-74435, Santa Cruz, Heidelberg, Germany), anti-A20 monoclonal antibody (sc-166692, Santa Cruz, Heidelberg, Germany), and anti-actin monoclonal antibody (MP6472J, MP Biomedicals, Eschwege, Germany).

Techniques: Activity Assay, Activation Assay, Mutagenesis, Gene Expression, Knockdown, Staining, Cell Analysis, Expressing, Western Blot, Inhibition

Journal: Virology Journal

Article Title: The expression and significance of proto-oncogene c-fos in viral myocarditis

doi: 10.1186/1743-422X-7-285

Figure Lengend Snippet: The expression change of c-fos oncogene mRNA in VMC mice

Article Snippet: c-fos monoclonal antibody, isoproterenol, normal goat serum, rabbit anti-c-fos oncogene protein, Biotinylated goat anti rabbit IgG, Streptavidin Biotin-peroxidase Complex (SABC), antigen restoration solution, pepsin, c-fos oligonucleotide probe, Occlusive solution, and rabbit anti digoxin were purchased from Boster Biological Technology Ltd.(Wuhan, China), Sigma Chemical Co. (Sigma, St.Louis, MO) and Biocompare Co.(South San Francisco, CA).

Techniques: Expressing, Control

Journal: Virology Journal

Article Title: The expression and significance of proto-oncogene c-fos in viral myocarditis

doi: 10.1186/1743-422X-7-285

Figure Lengend Snippet: The expression change of c-Fos oncogene protein in VMC mice

Article Snippet: c-fos monoclonal antibody, isoproterenol, normal goat serum, rabbit anti-c-fos oncogene protein, Biotinylated goat anti rabbit IgG, Streptavidin Biotin-peroxidase Complex (SABC), antigen restoration solution, pepsin, c-fos oligonucleotide probe, Occlusive solution, and rabbit anti digoxin were purchased from Boster Biological Technology Ltd.(Wuhan, China), Sigma Chemical Co. (Sigma, St.Louis, MO) and Biocompare Co.(South San Francisco, CA).

Techniques: Expressing, Control