Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Development and epigenetic plasticity of murine Müller glia

doi: 10.1016/j.bbamcr.2019.06.019

Figure Lengend Snippet: List of oligonucleotides and antibodies used in this study

Article Snippet: Isolation of Notch1+ cells, Glast+ cells, and cortical astrocytes We used the same protocol for immunomagnetic cell separation described in Dvoriantchikova et al. to isolate Notch1+ cells with monoclonal biotin-conjugated anti-Notch1 mouse antibodies (130–096-557, Miltenyi Biotec, Auburn, CA) [ 18 ].

Techniques:

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Development and epigenetic plasticity of murine Müller glia

doi: 10.1016/j.bbamcr.2019.06.019

Figure Lengend Snippet: Quantitative RT-PCR analysis has confirmed the microarray data for a group of selected genes. For each gene, the results are expressed as percentages ± SEM of the corresponding values in the Notch1+ cells isolated from P0 developing retinas (P0 Notch1+ cells).

Article Snippet: Isolation of Notch1+ cells, Glast+ cells, and cortical astrocytes We used the same protocol for immunomagnetic cell separation described in Dvoriantchikova et al. to isolate Notch1+ cells with monoclonal biotin-conjugated anti-Notch1 mouse antibodies (130–096-557, Miltenyi Biotec, Auburn, CA) [ 18 ].

Techniques: Quantitative RT-PCR, Microarray, Isolation

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Development and epigenetic plasticity of murine Müller glia

doi: 10.1016/j.bbamcr.2019.06.019

Figure Lengend Snippet: The results of the hierarchical and k-means clustering revealed the changes that are undergone in RPCs (Notch1+ cells) and Müller glia (Glast+ cells) during retinal development. A) Hierarchical clustering of differentially expressed genes showed that many P7 and P14 Notch1+ cells (late RPCs) are predisposed to become Müller glia. While Glast+ cells at P7 are still very close to a multipotent progenitor state, P14, P21, and P28 Glast+ cells are already grouped together indicating the adult Müller glia state. B) The 10 clusters were identified by the k-means clustering algorithm. C) The majority of differentially expressed genes were located in cluster 1, 3, and 4. D) Meanwhile, significant changes were undergone in genes located in small clusters 5, 7, and 8. A value F (F statistic) was calculated using the ANOVA test.

Article Snippet: Isolation of Notch1+ cells, Glast+ cells, and cortical astrocytes We used the same protocol for immunomagnetic cell separation described in Dvoriantchikova et al. to isolate Notch1+ cells with monoclonal biotin-conjugated anti-Notch1 mouse antibodies (130–096-557, Miltenyi Biotec, Auburn, CA) [ 18 ].

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Antiproliferative and Immunoregulatory Effects of Azelaic Acid Against Acute Myeloid Leukemia via the Activation of Notch Signaling Pathway

doi: 10.3389/fphar.2019.01396

Figure Lengend Snippet: Notch was found up-regulated after azelaic acid (AZA) treatment. (A) 120 differential expressed proteins associated in 52 functions after AZA treatment by protein mass spectrometry analysis in the heat map. (B) GO annotation with 528 differential expressed proteins. (C, D) THP-1, Molm-13, NK, and T cells were treated with 10 µM AZA for 24 h, the expression level of ICN1 and ICN2 was validated by western blot. (E) The expression of Notch1 and Notch2 in mouse spleen measured by ICH. IHC, immunohistochemistry.

Article Snippet: The following antibodies were used: CD3(Cat# 100203), CD4 (Cat# 100431), CD8 (Cat# 100761), CD107a (Cat# 328605), TRAIL (Cat# 308205), CD25 (Cat# 302605), and CD69 (Cat# 310903) were purchased from BioLegend (USA), ICN1 (Cat# CSB-PA084572), ICN2 (Cat# CSB-PA964902) were from CUSABIO (USA); GAPDH (Cat# 6004-1), β-actin (Cat# 14395-1) were from ProteinTech (USA).

Techniques: Mass Spectrometry, Expressing, Western Blot, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: Antiproliferative and Immunoregulatory Effects of Azelaic Acid Against Acute Myeloid Leukemia via the Activation of Notch Signaling Pathway

doi: 10.3389/fphar.2019.01396

Figure Lengend Snippet: Azelaic acid (AZA) exerts anti-leukemic effect by activating the Notch signaling pathway. (A) Notch responsive elements were transfected into 293T cells after 24 h. Cells were then treated with 10 µM AZA, 10 µM RO4929097, and combination for 24 h, the Notch signaling reporter assay was measured by dual luciferase reporter activity. (B) Validation of the RNA expression of Notch1 and Notch2, the downstream target genes HES1 and HEY1 in Molm-13 and THP-1 cells by qPCR. (C) The protein expression level of ICN1, ICN2, HEY1, and HES1 in acute myeloid leukemia (AML) cell lines after treatment of AZA and RO4929097 and their detection by western blot. ImageJ was used for the densitometric analysis. Data represent means ± SD. (D) Molm-13 cells were pretreated with 10 µM RO4939097 for 24 h, and then treated with 10 µM AZA for another 24 h. Cells were collected for apoptosis analysis. (E) NK cells and T cells were pretreated with 10 µM AZA, 10 µM RO4929097, and combination for 24h before co-culture with THP-1 cell and Molm-13 cells at an E:T ratio of 5:1. The cytotoxicity of NK and T was determined by detecting the LDH release rate. (F) NK and T cells were pre-treated with AZA and RO4929097, then co-cultured with THP-1 cell at an E:T ratio of 3:1 for 4 h. The level of TNF-α and IFN-γ in the supernatant was measured by ELISA. A Total of three independent experiments were performed. *P < 0.05, **P < 0.01,***P < 0.001.

Article Snippet: The following antibodies were used: CD3(Cat# 100203), CD4 (Cat# 100431), CD8 (Cat# 100761), CD107a (Cat# 328605), TRAIL (Cat# 308205), CD25 (Cat# 302605), and CD69 (Cat# 310903) were purchased from BioLegend (USA), ICN1 (Cat# CSB-PA084572), ICN2 (Cat# CSB-PA964902) were from CUSABIO (USA); GAPDH (Cat# 6004-1), β-actin (Cat# 14395-1) were from ProteinTech (USA).

Techniques: Transfection, Reporter Assay, Luciferase, Activity Assay, Biomarker Discovery, RNA Expression, Expressing, Western Blot, Co-Culture Assay, Cell Culture, Enzyme-linked Immunosorbent Assay