Fig. 6: LAMP3 is a downstream target gene of miR-423-3p that participates in the autophagy of NHAs. | Cell Death & Disease

Fig. 6: LAMP3 is a downstream target gene of miR-423-3p that participates in the autophagy of NHAs.

From: Inhibition of hypoxic exosomal miR-423-3p decreases glioma progression by restricting autophagy in astrocytes

Fig. 6

A, B NHAs were transfected with negative control mimics and miR-423-3p mimics. Utilizing a qPCR array, the heat map and volcano map showcased differential gene expression patterns. C, D The relative expression of Lamp3 was detected using western blotting in NHAs treated with PBS, U251-derived N-GDEs, H-GDEs, P3#GBM-derived N-GDEs, H-GDEs, control mimics, siNC, and miR-423-3p mimics; 3-MA and siBeclin1 were added to inhibit autophagy respectively. Below each blot, numbers represent the mean LAMP3 expression values from three independent experiments, quantified using grayscale analysis. Expression levels are normalized to a control set at 1. E A pulldown kit was used for NHAs to validate the connection between miR-423-3p and Lamp3; expression of human antigen R (HUR) was measured in negative and positive proteins (derived from the pulldown kit), and the NHA protein was added with biotin-marked sense and antisense RNA to confirm the successful pulldown of proteins. The expression of LAMP3 and GAPDH was detected in NHA protein treated with biotin-marked sense and antisense RNA. F A schematic diagram was created using biorender.com. GBM glioblastoma multiforme, H-GDE hypoxic glioma-derived exosome, N-GDE normoxic glioma-derived exosome, NHA normal human astrocyte.

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