Journal: Cell Death & Disease

Article Title: Gadd45a opens up the promoter regions of miR-295 facilitating pluripotency induction

doi: 10.1038/cddis.2017.497

Figure Lengend Snippet: Gadd45a upregulates miR-290 clusters in reprogramming. ( a ) Phase contrast and fluorescence photographs of MEFs infected with SKO plus Flag or Gadd45a, SKOM plus Flag or Gadd45a. Flag is the empty vector control. Arrows point to emerging colonies. ( b ) Gadd45a significantly enhances iPSC generation in various mediums. In the absence of vitamin C, the numbers of GFP+ colonies were counted at day 17; in the presence of vitamin C, the numbers of GFP+ colonies were counted at day 13; in iCD1 medium, the numbers of GFP+ colonies were counted at day 7. Data are represented as mean±S.D.; n =3. * P ≤0.05, ** P ≤0.01, *** P ≤0.001. ( c ) Gadd45a overexpression blocks the formation of partially reprogrammed cells. The colonies were stained for alkaline phosphatase (AP). The numbers of AP+GFP+ colonies and AP+GFP− colonies were counted. Data are represented as mean±S.D. n =3; *** P ≤0.001. ( d ) Heatmaps depicting the relative fold change of miRNA expression at day 8 post infection by miRNA microarray. Red and green colors indicated increased and decreased expression, respectively. ( e ) qPCR analysis of miR-294 and miR-295 expression level during SKO- and SKOM-mediated reprogramming in the presence and absence of Gadd45a. Data are represented as mean±S.D.; n =3. *** P ≤0.001

Article Snippet: Real-time RT-PCR was performed with the All-in-One miRNA qRT-PCR Detection Kit (GeneCopoeia, Rockville, MD, USA) following the manufacturer’s instructions.

Techniques: Fluorescence, Infection, Plasmid Preparation, Control, Over Expression, Staining, Expressing, Microarray

Journal: Frontiers in Immunology

Article Title: Experimental Demyelination and Axonal Loss Are Reduced in MicroRNA-146a Deficient Mice

doi: 10.3389/fimmu.2018.00490

Figure Lengend Snippet: Differential expression of microRNAs (miRNAs) in the corpus callosum of mice during experimental demyelination and remyelination. Using microarray and validation by quantitative PCR (qPCR), three miRNAs, miR-146a (A) , miR-193a (B) and miR-181b (C) were differentially regulated in response to CPZ exposure in the corpus callosum. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 5–8 in each group, one-way ANOVA, mean ± SEM. Abbreviation: C: un-manipulated controls covering ages of mice used in the de- and remyelinating experiments, 4wD: 4 weeks demyelination. 2dR: 2 days (acute) remyelination, 2wR: 2 weeks (full) remyelination.

Article Snippet: For microRNA profiling, the Agilent Mouse miRNA Microarray Kit (G4472A, 8 × 15k) was applied according to the manufacturer’s instruction (version 1.0) with 100–100 ng quality-checked total RNA.

Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Experimental Demyelination and Axonal Loss Are Reduced in MicroRNA-146a Deficient Mice

doi: 10.3389/fimmu.2018.00490

Figure Lengend Snippet: Expression of experimentally validated miR-146a target genes and proteins in response to CPZ exposure in wild-type and miR-146-deficient mice. (A) Expression of miR-146a target genes (miRTarBase) among 1,239 differentially expressed genes in the corpus callosum in response to CPZ exposure determined by a 4 × 44K Agilent Whole Mouse Genome Microarray (NCBI Gene Expression Omnibus with accession GSE100663). (B) The protein expression of validated miR-146a target genes SMAD4 and SNAP25 were downregulated in the corpus callosum of miR-146a KO mice compared to WT mice by liquid chromatography mass spectrometry in response to CPZ. (C) SNAP25 and (D) SMAD4 in corpus callosum lysates of miR-146a KO and WT mice in response to CPZ examined by ELISA. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 8–4 in each group, mean ± SEM. Abbreviations: C: un-manipulated controls, 4wD: 4 weeks demyelination. 2dR: 2 days (acute) remyelination, 2wR: 2 weeks (full) remyelination.

Article Snippet: For microRNA profiling, the Agilent Mouse miRNA Microarray Kit (G4472A, 8 × 15k) was applied according to the manufacturer’s instruction (version 1.0) with 100–100 ng quality-checked total RNA.

Techniques: Expressing, Microarray, Liquid Chromatography, Mass Spectrometry, Enzyme-linked Immunosorbent Assay