Fig. 3.

Knockdown of NEAT1 affects signaling cascades related to autophagy in primary microglia exposed to hypoxia. a–d Autophagy-related genes, i.e., Atg3 (a), Atg5 (b), Beclin1 (c), and STAT3 (d) were determined by quantitative qRT-PCR normalized to PPIA. e–h NEAT1 knockdown repressed LC3 but activated p62 expressions. Western blot was utilized to analyze the expression of LC3 (e) at a 17 kDa band and p62 (f) at a 62 kDa band in microglial cells in four parallel groups (normoxia, OGD, OGD + ASO scramble, and OGD + ASO NEAT1). The relative quantitative analysis was normalized against β-actin or GAPDH. Results are expressed as mean ± standard deviation and analyzed by one-way ANOVA followed by Tukey's post-hoc-test. NS, no significance, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Abbreviations: NEAT1, nuclear paraspeckle assembly transcript 1; ASO, antisense oligonucleotide; OGD, oxygen-glucose-deprivation; Atg3, autophagy-related 3; PPIA, peptidylprolyl isomerase A