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HLCs were cultured with Alexa-Flour 488-ac-LDL for 1 hour, followed by immunohistochemistry and DAPI counterstaining [1]
Single-cell suspensions were then loaded onto microfluidic devices, and single-cell RNA sequencing (scRNA-seq) libraries were constructed according to the Singleron GEXSCOPE protocol via the GEXSCOPE Single-Cell RNA Library Kit (Singleron), which included cell disruption, mRNA trapping, cell labeling (with barcodes) and mRNA labeling (with unique molecular identifier (UMI)), reverse transcription of mRNA into complementary DNA (cDNA) and amplification, cDNA fragmentation, and finally cDNA fragment cloning
15LO1 increases intracellular oxidative potential by lowering GSH and increasing extracellular GSSG To confirm that 15LO1-PEBP1 complex activity alters IL-13induced intracellular GSH levels, we modeled these relationships in vitro by stimulating cultured HAECs (at ALI) with IL-13 for 710 days in the presence and absence of 15LO1 protein (dicer siRNA knockdown [KD] of ALOX15 gene) or enzymatic activity (specific 15LO1 inhibitor BLX2477) (14, 15)
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Subsequently, the slides were placed into a humid box and were then incubated with Rabbit anti-LC3B (1:500 dilutions, Abcam) primary antibody
