ipsc line from fibroblast control #1  (Coriell Institute for Medical Research)

Journal: Scientific Reports

Article Title: Functional roles of pantothenic acid, riboflavin, thiamine, and choline in adipocyte browning in chemically induced human brown adipocytes

doi: 10.1038/s41598-024-69364-w

Figure Lengend Snippet: Concentration-dependent effects of PA on mitochondrial respiration and glycolysis. ( A ) Oxygen consumption rate (OCR) was measured using Seahorse XFe96 extracellular flux analyzer in control fibroblasts, NoC(V7) (grey circles), and ciBAs converted in the absence of PA, RoFB(V7) (black diamonds). Mitochondrial respiration inhibitors, oligomycin, FCCP, and antimycin A/rotenone, were added during the measurement, as indicated. P values were determined using two-way ANOVA. ( B ) OCR was compared between RoFB(V7) (grey circles) and either RoFB(V7 + PA, 0.5 μg/mL), RoFB(V7 + PA, 4 μg/mL), or RoFB(V7 + PA, 16 μg/mL) (black diamonds). P values were determined using two-way ANOVA. ( C,D ) OCR corresponding to basal respiration, maximal respiration, ATP production, and proton leak was compared. ( E ) Extracellular acidification rate (ECAR) was measured in control fibroblasts, NoC(V7) (grey circles), and ciBAs in the absence of PA, RoFB(V7) (black diamonds). Glucose, oligomycin, and 2-deoxyglucose (2-DG) were sequentially added during the measurement, as indicated. P values were determined using two-way ANOVA. ( F ) ECAR was also compared between the control ciBAs, RoFB(V7) (grey circles), and either RoFB(V7 + PA, 0.5 μg/mL), RoFB(V7 + PA, 4 μg/mL), or RoFB(V7 + PA, 16 μg/mL) (black diamonds). P values were determined using two-way ANOVA. ( G ) ECAR corresponding to glycolysis and glycolytic capacity was calculated. Data represent mean ± SD ( n = 6–8). ( H ) Lactate secretion into culture supernatants was quantified in control fibroblasts and ciBAs treated with PA at various concentrations throughout the experiments. Data represent mean ± SD ( n = 3). ( I ) Mitochondrial membrane potential (MMP) was evaluated by staining with the fluorescent probe, MT-1 dye. The area of the staining for the dye was quantified by ImageJ software. Data represent mean ± SD ( n = 5). One-way ANOVA with Tukey’s multiple comparison tests: * p < 0.05, ** p < 0.01, *** p < 0.001, N.S.; not significant.

Article Snippet: In brief, the cells were pre-incubated with MitoTracker for 30 min at 37 °C in a 5% CO 2 before the cells were fixed with 4% paraformaldehyde for 10 min. MMP in control fibroblasts and ciBAs were stained using the MT-1 MitoMP Detection kit (MT13, Dojindo).

Techniques: Concentration Assay, Control, Membrane, Staining, Software, Comparison

Journal: Scientific Reports

Article Title: Functional roles of pantothenic acid, riboflavin, thiamine, and choline in adipocyte browning in chemically induced human brown adipocytes

doi: 10.1038/s41598-024-69364-w

Figure Lengend Snippet: Comparison of the transcriptome in ciBAs treated with PA at low and high concentrations throughout the conversion. ( A ) Multidimensional scaling analysis graphically indicates the similarity and variability of the transcriptome in the control fibroblast, NoC(V7), ciBAs, RoFB(V7), and ciBAs treated with PA at low and high concentrations, RoFB(V7 + PA, 0.5 μg/mL) and RoFB(V7 + PA, 16 μg/mL). ( B ) Gene ontology (GO) enrichment analysis was performed in upregulated differentially expressed genes (DEGs) in RoFB(V7 + PA, 0.5 μg/mL) compared with RoFB(V7). The top 10 GO terms are represented in the category of biological process. ( C ) GO analysis was performed in downregulated DEGs in RoFB(V7 + PA, 16 μg/mL) compared with RoFB(V7 + PA, 0.5 μg/mL). ( D ) The FPKM values in the RNA-Seq results indicate the transcriptional levels of UCP1 , CKMT1A , CKMT1B , CKMT2 , CKB , ALPL ( TNAP ), SLC6A8 , and CKM genes. ( E ) UCP1 , CKMT1 , CKMT2 , CKB , ALPL ( TNAP ), and SLC6A8 mRNA were measured by qRT-PCR analysis in ciBAs treated with PA as indicated. Data represent mean ± SD (n = 3). One-way ANOVA with Tukey’s multiple comparison tests: * P < 0.05, ** P < 0.01, *** P < 0.001, N.S.; not significant.

Article Snippet: In brief, the cells were pre-incubated with MitoTracker for 30 min at 37 °C in a 5% CO 2 before the cells were fixed with 4% paraformaldehyde for 10 min. MMP in control fibroblasts and ciBAs were stained using the MT-1 MitoMP Detection kit (MT13, Dojindo).

Techniques: Comparison, Control, RNA Sequencing, Quantitative RT-PCR

Journal: Neuron

Article Title: Kir4.1-Dependent Astrocyte-Fast Motor Neuron Interactions Are Required for Peak Strength

doi: 10.1016/j.neuron.2018.03.010

Figure Lengend Snippet:

Article Snippet: Human: iPSC line from Fibroblast Control #1 , Coriell Institute , Coriell ND41866 ∗ C.

Techniques: Virus, Plasmid Preparation, Recombinant, SYBR Green Assay, Control, Mutagenesis, Software