Journal: PLoS ONE

Article Title: Functional Profiling Reveals Critical Role for miRNA in Differentiation of Human Mesenchymal Stem Cells

doi: 10.1371/journal.pone.0005605

Figure Lengend Snippet: (A–G) Transfection with inhibitors of miR-489 and -27a results in up-regulation of GCA protein expression. (A–F). hMSC were transfected with control molecule, ic1 (25 nM) or with combination of inhibitors, i489+i27a (12.5 nM each). GCA protein expression was determined by immunofluorescence as described in . Cells were stained with Hoechst 33342 (A–C) and GCA specific primary antibody, followed by a secondary antibody (E,F). Control wells were treated with secondary antibody only and demonstrate no specific staining (D). (G) Quantitation of GCA immunohystochemistry. Data shown represents three independent transfections, four fields each. (mean+/−SD). ** - Student's ttest p value between treated cells and corresponding control group, p<0.05. (H) RT-PCR-based study of gene expression demonstrated that the expression of GCA mRNA is regulated by miR-489 and -27a. hMSC were transfected with Inhibitor Control molecule 1 (IC1, 25 nM) or with a combination of inhibitors for miR-27 and -489 (i27a+i489, 12.5 nM each). Transfected cells were incubated for 2 days in propagation media and then harvested for totat RNA. (I) siRNA-mediated knockdown of GCA (grancalcin) resulted in a significant decrease of AP activity in hMSC under differentiation conditions. hMSC were transfected with Control siRNA, RUNX2 and GCA siRNA (each at 50 nM). Cells were switched to osteogenic media 24 hr after transfection and harvested 6 days after the switch. Data shown represents two independent transfection experiments performed in duplicate (H) or triplicate (I) (mean+/−SD). **: Student's ttest p value between treated cells and corresponding control group, p<0.05.

Article Snippet: Corresponding TaqMan® Gene Expression Assays were purchased from Life Technologies (Carlsbad, CA) - GCA Assay ID Hs00201854_m1, SLC22A2 Assay ID Hs00533907_m1, PEX7 Assay ID Hs00165464_m1, AHSG Assay ID Hs00155659_m1, and CHRD Assay ID Hs00415315_m1.

Techniques: Transfection, Expressing, Control, Immunofluorescence, Staining, Quantitation Assay, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Incubation, Knockdown, Activity Assay

Journal: BMC Medicine

Article Title: Diagnostic delay for giant cell arteritis – a systematic review and meta-analysis

doi: 10.1186/s12916-017-0871-z

Figure Lengend Snippet: Characteristics of articles reporting delay of giant cell arteritis (GCA) diagnosis

Article Snippet: Nesher [ ] , GCA defined using 1990 ACR criteria c , 144 , 64.6 , 73.0 , – , – , M , 1.5 , – , 0.1–12 , 6.4 , 7.9.

Techniques: Sampling, Biomarker Discovery

Journal: BMC Medicine

Article Title: Diagnostic delay for giant cell arteritis – a systematic review and meta-analysis

doi: 10.1186/s12916-017-0871-z

Figure Lengend Snippet: Extent of diagnostic delay reported within articles included in systematic review ( n = 22)

Article Snippet: Nesher [ ] , GCA defined using 1990 ACR criteria c , 144 , 64.6 , 73.0 , – , – , M , 1.5 , – , 0.1–12 , 6.4 , 7.9.

Techniques: Diagnostic Assay, Biomarker Discovery, Sedimentation