Fig. 2.

Knockdown of NEAT1 affects expression patterns of LD in primary microglia exposed to hypoxia. a The fluorescence staining of neutral LD (BODIPY, green) in microglial cells (CD11b, red) cultured with ASO scramble or ASO NEAT1. b Statistical analysis of the percentage of LD in whole cells. c, d Primary microglia were transfected with ASO NEAT1 (or ASO scramble as control) for 24 h to generate a stable NEAT1 knockdown. The expression of the LD-related genes PLIN2 (c) and TREM2 (d) was detected using qRT-PCR in comparison to PPIA. e–g Western blot analysis of PLIN2 and TREM2 expression in OGD/R microglial cells after treatment with ASO scramble or ASO NEAT1. The relative quantitative analysis was normalized against GAPDH or tublin, as indicated. All 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/R, oxygen-glucose-deprivation/reoxygenation; LD, lipid droplets; PLIN2, perilipin 2; TREM2, triggering receptor expressed on myeloid cells 2; PPIA, peptidylprolyl isomerase A