Figures & data
Figure 1. Inflammation and NEAT1 expression are promoted in the LPS-evoked cells. (a) Cell viability and (b and c) apoptosis rate of the PC-12 cell induced by 0, 2, 4, 8, 16 μg/ml LPS. (e) Expression levels of NEAT1 in the cells incubated with 0, 2, 4, 8, 16 μg/ml LPS. Three independent experiments were carried out. *P < 0.05, vs. 0 μg/ml LPS; **P < 0.01, vs. 0 μg/ml LPS. LPS, lipopolysaccharide; NEAT1, nuclear enriched abundant transcript 1
Figure 2. Knockdown of NEAT1 enhances cell viability, and restrains cell apoptosis and inflammation. (a) Si-NEAT1 was successfully synthesized and transfected into the cells. (b) Cell viability and (c and d) apoptosis of the control group and the LPS-evoked cells transfected with si-NEAT1. (e) The protein expressions of bax, cleaved caspase-3 and bcl-2. (f) mRNA expressions of IL-1β, IL-6, IL-10, and TNF-α in the control group and the LPS-evoked cells transfected with si-NEAT1. Three independent experiments were carried out. **P < 0.01, vs. control or LPS+si-nc. LPS, lipopolysaccharide; si, small interference RNA; NEAT1, nuclear enriched abundant transcript 1; nc, negative control
Figure 3. NEAT1 directly targets miR-211-5p. (a) Wild and mutant type of NEAT1 luciferase reporters were constructed and co-transfected with miR-211-5p into the cells. (b) Luciferase activities of the wild and mutant groups. (c) RT-qPCR was used to detect the enrichment of NEAT1 in miR-211-5p biotin-labeled group and control group. (d) MiR-211-5p expression level of the cells transfected with si-NEAT1. (e) MiR-211-5p levels of the cells incubated with different concentrations of LPS were detected. Three independent experiments were carried out. **P < 0.01, vs. mimic NC, control probe, si-nc or 0 μg/ml LPS. LPS, lipopolysaccharide; si, small interference RNA; NEAT1, nuclear enriched abundant transcript 1; wt, wild type; mut, mutant type; nc, negative control
Figure 4. Inhibition of miR-211-5p antagonizes the effects of NEAT1 knockdown on cell viability, apoptosis and inflammation. (a) MiR-211-5p inhibitor was successfully constructed and transfected into the cells. (b) Cell viability and (c and d) apoptosis of the control group and the LPS-evoked cells co-transfected with si-NEAT1 and miR-211-5p inhibitor. (e) The protein expressions of bax, cleaved caspase-3 and bcl-2. (f) mRNA expressions of IL-1β, IL-6, IL-10, and TNF-α in the control group and the LPS-evoked cells co-transfected with si-NEAT1 and miR-211-5p inhibitor. Three independent experiments were carried out. *P < 0.05, vs. control; **P < 0.01, vs. control; #P < 0.05, vs. LPS+si-nc; &P < 0.05, vs. LPS+si-NEAT1+ inh-nc. LPS, lipopolysaccharide; si, small interference RNA; NEAT1, nuclear enriched abundant transcript 1; wt, wild type; mut, mutant type; nc, negative control
Figure 5. MAPK1 is the downstream target gene of miR-211-5p. (a) Wild and mutant type of MAPK1 luciferase reporters were constructed and co-transfected with miR-211-5p into the cells. (b) Luciferase activities of the wild and mutant groups. (c) RT-qPCR was used to detect the enrichment of MAPK1 in miR-211-5p biotin-labeled group and control group. (d) MRNA expression levels of MAPK1 were analyzed in the miR-211-5p or miR-211-5p inhibitor transfected group. (e) MAPK1 levels of the cells incubated with different concentrations of LPS were detected. Three independent experiments were carried out. **P < 0.01, vs. miR-nc or biotin-nc; #P < 0.05, vs. inhibitor-nc; ##P < 0.01, vs. inhibitor-nc. MAPK1, mitogen-activated protein kinase 1; wt, wild type; mut, mutant type; nc, negative control
Figure 6. Overexpression of MAPK1 antagonizes the effects of NEAT1 knockdown. (a) MAPK1 overexpression vector was successfully constructed and transfected into the cells. (b) Cell viability and (c and d) apoptosis of the control group and the LPS-evoked cells co-transfected with si-NEAT1 and MAPK1. (e) The protein expressions of bax, cleaved caspase-3 and bcl-2 in the cells co-transfected with si-NEAT1 and MAPK1. (f) mRNA expressions of IL-1β, IL-6, IL-10, and TNF-α in the control group and the LPS-evoked cells co-transfected with si-NEAT1 and MAPK1. Three independent experiments were carried out. *P < 0.05, vs. control; **P < 0.01, vs. control; #P < 0.05, vs. LPS+si-nc; &P < 0.05, vs. LPS+si-NEAT1+ inh-nc. LPS, lipopolysaccharide; si, small interference RNA; NEAT1, nuclear enriched abundant transcript 1; wt, wild type; mut, mutant type; nc, negative control
