Journal: Toxins

Article Title: Identification of Potential Therapeutic Targets Against Anthrax-Toxin-Induced Liver and Heart Damage.

doi: 10.3390/toxins17020054

Figure Lengend Snippet: Figure 3. Identification of EdTx-induced, cytotoxicity-related genes. Total RNA was isolated from primary hepatocytes treated with PBS or EdTx (4 µg/mL) for 6 h, and the samples were subjected to microarray analysis using the GeneChips Mouse Transcriptome Assay 1.0. Some genes (218) were found to have significant expression changes in primary hepatocytes exposed to EdTx treatment com- pared with PBS-treated cells. The Partek Genomics Suite was used to analyze the signaling pathways associated with these differentially expressed genes. (A) The pathways with enrichment score > 2 and p-value < 0.05 are shown. The numbers at the top of each column show the number of genes that have expression changes in each pathway. (B) The 35 significantly changed genes and sequencing expression fold changes. * p < 0.05, ** p < 0.01 vs. PBS. (C) The results of a protein–protein network analysis among the 35 genes are shown. Nine genes that are involved in glycogen metabolism, cAMP production, and cell apoptosis are marked with red circles and were further investigated in the following experiments. (D) These potential EdTx-induced cytotoxicity-related genes were knocked down individually or in combination in primary hepatocytes using the corresponding siRNAs. si-CMG2 was used as a positive control, and si-GFP and si-(no gene) were used as negative controls. Primary hepatocytes deficient in these genes were treated with PBS or EdTx (4 µg/mL) for 6 h. The intracellular concentration of cAMP was determined using ELISA. * p < 0.05, ** p < 0.01 vs. si-GFP (n = 3). 4mix, Ramp3 + Rgs1 + Pck1 + G6pc; 5mix, Hcar2 + Fosl2 + Fos + Cxcl2 + Cxcl3.

Article Snippet: siRNAs targeting the murine Ramp3 (si-Ramp3), Rgs1 (si-Rgs1), Pck1 (si-Pck1), G6pc (si-G6pc), Hcar2 (si-Hcar2), Fosl2 (si-Fosl2), Fos (si-Fos), Cxcl2 (si-Cxcl2), Cxcl3 (si-Cxcl3), and Cmg2 (si-CMG2) genes were provided by Santa Cruz Biotechnology.

Techniques: Isolation, Microarray, Expressing, Protein-Protein interactions, Sequencing, Positive Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Toxins

Article Title: Identification of Potential Therapeutic Targets Against Anthrax-Toxin-Induced Liver and Heart Damage.

doi: 10.3390/toxins17020054

Figure Lengend Snippet: Figure 3. Identification of EdTx-induced, cytotoxicity-related genes. Total RNA was isolated from primary hepatocytes treated with PBS or EdTx (4 µg/mL) for 6 h, and the samples were subjected to microarray analysis using the GeneChips Mouse Transcriptome Assay 1.0. Some genes (218) were found to have significant expression changes in primary hepatocytes exposed to EdTx treatment com- pared with PBS-treated cells. The Partek Genomics Suite was used to analyze the signaling pathways associated with these differentially expressed genes. (A) The pathways with enrichment score > 2 and p-value < 0.05 are shown. The numbers at the top of each column show the number of genes that have expression changes in each pathway. (B) The 35 significantly changed genes and sequencing expression fold changes. * p < 0.05, ** p < 0.01 vs. PBS. (C) The results of a protein–protein network analysis among the 35 genes are shown. Nine genes that are involved in glycogen metabolism, cAMP production, and cell apoptosis are marked with red circles and were further investigated in the following experiments. (D) These potential EdTx-induced cytotoxicity-related genes were knocked down individually or in combination in primary hepatocytes using the corresponding siRNAs. si-CMG2 was used as a positive control, and si-GFP and si-(no gene) were used as negative controls. Primary hepatocytes deficient in these genes were treated with PBS or EdTx (4 µg/mL) for 6 h. The intracellular concentration of cAMP was determined using ELISA. * p < 0.05, ** p < 0.01 vs. si-GFP (n = 3). 4mix, Ramp3 + Rgs1 + Pck1 + G6pc; 5mix, Hcar2 + Fosl2 + Fos + Cxcl2 + Cxcl3.

Article Snippet: siRNAs targeting the murine Ramp3 (si-Ramp3), Rgs1 (si-Rgs1), Pck1 (si-Pck1), G6pc (si-G6pc), Hcar2 (si-Hcar2), Fosl2 (si-Fosl2), Fos (si-Fos), Cxcl2 (si-Cxcl2), Cxcl3 (si-Cxcl3), and Cmg2 (si-CMG2) genes were provided by Santa Cruz Biotechnology.

Techniques: Isolation, Microarray, Expressing, Protein-Protein interactions, Sequencing, Positive Control, Concentration Assay, Enzyme-linked Immunosorbent Assay