Research Paper Volume 12, Issue 9 pp 7660—7678

HOTAIR drives autophagy in midbrain dopaminergic neurons in the substantia nigra compacta in a mouse model of Parkinson’s disease by elevating NPTX2 via miR-221-3p binding

Figure 2. HOTAIR depletion represses autophagy in dopaminergic neurons in SNc tissues of PD mice. The expression patterns of HOTAIR in SNc tissues measured by RT-qPCR (n = 10), *p < 0.05 vs. mice injected with saline; #p < 0.05 vs. mice injected with MPTP + LV-sh-NC. (A) The ratios of LC3B-I/LC3B-II and LAMP1/LAMP2 along with the P62 expression patterns in SNc tissues determined by Western blot analysis (n = 10), *p < 0.05 vs. mice injected with saline; #p < 0.05 vs. mice injected with MPTP + LV-sh-NC. (B) The expression patterns of HOTAIR in MN9D cells treated with PBS, MPP+, MPP+ + sh-NC and MPP+ + sh-HOTAIR measured by RT-qPCR, *p < 0.05 vs. MN9D cells treated with PBS; #p < 0.05 vs. MN9D cells treated with MPP+ + sh-NC. (C) The ratios of LC3B-I/LC3B-II and LAMP1/LAMP2 along with the P62 expression patterns in MN9D cells treated with PBS, RAPA, MPP+, MPP+ + sh-NC and MPP+ + sh-HOTAIR assessed by Western blot analysis, *p < 0.05 vs. MN9D cells treated with PBS; #p < 0.05 vs. MN9D cells treated with MPP+ + sh-NC. (D) Detection of autophagy levels in MN9D cells by co-localization analysis. (E) Data (mean ± standard deviation) among multiple groups were analyzed using one-way ANOVA and subjected to Tukey’s post-hoc test. The experiment was repeated three times. HOTAIR, HOX transcript antisense intergenic RNA; SNc, substantia nigra compact; PD, Parkinson’s disease; RT-qPCR, reverse transcription quantitative polymerase chain reaction; PBS, phosphate buffered saline; MPP+, 1-methyl-4-phenylpyridinium; NC, negative control; ANOVA, analysis of variance; MPTP, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyrindine.