Human lncrna microarray version 3

Peripheral blood RNAs were extracted using a QIAamp RNA Blood Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's guidelines. Blood RNAs from HC (n = 4), ED without liver disease (n = 4), and AC (AC1 from Child‐Pugh class A, n = 4; AC2 from Child‐Pugh class B, n = 4; and AC3 from Child‐Pugh class C, n = 4) were subjected to global transcriptomic profiling using the Arraystar human lncRNA microarray version 3.0 (Arraystar, Rockville, MD). Quantitative polymerase chain reaction (qPCR) was used to validate the expression of the top 20 peripheral blood lncRNAs in a separate cohort of 17 HC, 19 ED, and 48 AC subjects as well as in the liver tissues from controls (n = 24) and subjects with AC (n = 19).

Two groups of cells (hMSCs from the same patient undergoing expansion and chondrogenic induction to chondrogenesis), which each contained the mixture of cell sources from three parallel experiment, were prepared, and total RNA was isolated using TRIzol reagent. RNA quality and quantity were measured by Nanodrop 2000 (Thermo Fisher Scientific). RNA integrity was assessed by standard denaturing agarose gel electrophoresis. For microarray analysis, Agilent Array platform was employed. The sample preparation and microarray hybridization were performed based on the manufacturer’s standard protocols with minor modifications. All labeled lncRNAs were hybridized onto an Arraystar Human LncRNA Microarray version -3.0 (Arraystar, Kangcheng, Shanghai, China). The microarray screening data have been submitted to the GEO database (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE128533). A heatmap was exported using Agilent Feature Extraction software (version 11.0.11, Agilent Technologies). Differentially expressed lncRNAs with statistical significance between the two samples were identified through fold change filtering.