Journal: Physiological Reports

Article Title: CXCL10 is a novel anti‐angiogenic factor downstream of p53 in cardiomyocytes

doi: 10.14814/phy2.15304

Figure Lengend Snippet: p53‐induced CXCL10, secreted from cardiomyocytes, inhibited the tube formation of endothelial cells. Culture media were collected from p53‐ or β‐gal‐overexpressing NRCMs. RAOECs were cultured with the conditioned media in the presence of AMG487, an inhibitor of CXCR3. Angiogenesis assay was performed. (a) Representative images were shown. (b) The number of tube formation was measured ( n = 6). * p < 0.05, ** p < 0.01 by one‐way ANOVA followed by Tukey–Kramer test.

Article Snippet: After coating the wells with the extracellular matrix, 5 × 10 5 RAOECs were seeded to each well and cultured with the conditioned media contained AMG 487 (MedChemExpress), a selective antagonist of CXCR3.

Techniques: Cell Culture, Angiogenesis Assay

Journal: EBioMedicine

Article Title: A Minimally-invasive Blood-derived Biomarker of Oligodendrocyte Cell-loss in Multiple Sclerosis

doi: 10.1016/j.ebiom.2016.06.031

Figure Lengend Snippet: Murine brain and spinal cord show differential methylation in the MOG gene, due to their O4 + cell population. A. Sanger sequencing results of bisulfite treated DNA from murine tissues. Arrows point toward CpG sites where cytosines (C) are preserved in methylated samples (Liver, Kidney), or converted to thymines (T) in samples containing demethylated CpGs, leading to a mixed population of C′s and T's (Brain, Spinal Cord). B. Methylation-sensitive DNA digestion was performed on the magnetically spinal cord of liver derived-DNA using the McrBC enzyme. Digested DNA was subjected to semi-quantitative PCR (cycles 32 and 35) and run on agarose gel. UnTx—untreated DNA receiving all reaction components except McrBC enzyme. Pos—positive control using cloned native MOG DNA. NTC—no template control. C and D. O4 + and O4 − cells were separated from four separate murine brains by magnetic beads. FACS analysis showed > 92.6 ± 3.9% enrichment of O4 + cells among four independent preparations when compared to O4 − fractions. E. DNA from murine O4 + cells is differentially methylated in the MOG gene compared to DNA from O4 − cells, the SW10 Schwann cell line, and liver. Sequence analysis was performed on first-step PCR product of each sample, 10 clones from each are shown (○ represent demethylated cytosines; ●, methylated cytosines). Locations in relation to the MOG transcription start site are listed, methylation-specific murine primers incorporate the CpG site at bp + 2752.

Article Snippet: SW10 immortalized neuronal murine Schwann cells were purchased from American Type Culture Collection (Manassas, VA) and cultured using the provided protocols.

Techniques: Methylation, Sequencing, Derivative Assay, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Positive Control, Clone Assay, Control, Magnetic Beads

Journal: EBioMedicine

Article Title: A Minimally-invasive Blood-derived Biomarker of Oligodendrocyte Cell-loss in Multiple Sclerosis

doi: 10.1016/j.ebiom.2016.06.031

Figure Lengend Snippet: Methylation-specific primers display high specificity and sensitivity and can detect demethylated MOG-DNA in murine brain, spinal cord, and O4 + cells. A. A depiction of MOG gene region utilized for mouse methylation-specific qPCR analysis; cytosine at bp + 2752 from MOG transcription start site incorporated into reverse primer sequence. B. Methylation-specific primers tested using plasmids containing methylated and demethylated murine MOG-DNA inserts over a wide range of serial dilutions (R 2 = 0.987, p < 0.0001). C. Methylation-specific primers were used in qPCR with bisulfite treated DNA from murine liver, kidney, brain, and spinal cord. Three independent analyses used to compute DMI averages; Liver vs. Brain/Spinal Cord p < 0.001, Kidney vs. Brain/Spinal Cord p < 0.001. D. Methylation-specific primers were used in qPCR with murine O4 + cells, O4 − cells, and SW10 Schwann cells. Three independent analyses used to compute DMI averages; ANOVA p = 0.0008, O4 + vs. O4 − p < 0.01, O4 + vs. SW10 Schwann cells p < 0.01.

Article Snippet: SW10 immortalized neuronal murine Schwann cells were purchased from American Type Culture Collection (Manassas, VA) and cultured using the provided protocols.

Techniques: Methylation, Sequencing