Journal: Journal of translational medicine

Article Title: FTO-targeted siRNA delivery by MSC-derived exosomes synergistically alleviates dopaminergic neuronal death in Parkinson's disease via m6A-dependent regulation of ATM mRNA.

doi: 10.1186/s12967-023-04461-4

Figure Lengend Snippet: Fig. 4 FTO promotes ATM expression through m6A-dependent stabilizing of ATM mRNA in dopaminergic neurons. A The potential m6A sites on ATM mRNA were analyzed by m6Avar. B qRT-PCR analysis of the indicated genes in MN9D dopaminergic neurons transfected with si-NC or si-FTO (n = 3). C qRT-PCR analysis of the indicated genes in MN9D dopaminergic neurons transfected with the empty control plasmid or pCMV-Flag-FTO plasmid (n = 3). D qRT-PCR analysis of ATM mRNA in MN9D dopaminergic neurons transfected with pCMV-Flag-FTO plasmid or treated with MPP + (n = 3). E qRT-PCR analysis of ATM mRNA in MN9D dopaminergic neurons co-transfected with pCMV-Flag-FTO plasmid + pCMV-Myc vector or pCMV-Flag-FTO plasmid + pCMV-Myc-Mettl3 (n = 3). Data are represented as mean ± SD, *represents P < 0.05, **represents P < 0.01

Article Snippet: The primary antibodies, including alpha synuclein antibody (α-syn, 10842-1-AP, Proteintech, China), TH antibody (25859-1-AP, Proteintech, China), FTO antibody (27226-1-AP, Proteintech, China), and ATM antibody (27156-1-AP, Proteintech, China) were used.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Plasmid Preparation

Journal: Journal of translational medicine

Article Title: FTO-targeted siRNA delivery by MSC-derived exosomes synergistically alleviates dopaminergic neuronal death in Parkinson's disease via m6A-dependent regulation of ATM mRNA.

doi: 10.1186/s12967-023-04461-4

Figure Lengend Snippet: Fig. 6 Synergistic si-FTO loading of exosomes effectively protect dopaminergic neuronal death in PD model in vivo. A Confocal microscopy analysis of the distributions of MSC-exo (red)-si-FTO (green) in the striatum of mice PD model. B and C qRT-PCR analysis of FTO and ATM mRNA expressions in the striatum of mice PD model injected with MSC-exo-si-NC or MSC-exo-si-FTO (n = 3). D Western blotting analysis of TH, α-Syn, ATM and FTO protein expressions in the striatum of mice PD model injected with MSC-exo-si-NC or MSC-exo-si-FTO. E Immunofluorescence staining analysis of FTO and TH expressions in the striatum of mice PD model injected with MSC-exo-si-NC or MSC-exo-si-FTO (n = 3). F Immunofluorescence staining analysis of FTO and ATM expressions in the striatum of mice PD model injected with MSC-exo-si-NC or MSC-exo-si-FTO (n = 3). G Immunofluorescence staining analysis of the apoptosis of dopaminergic neurons in the striatum of mice PD model injected with MSC-exo-si-NC or MSC-exo-si-FTO (n = 3).Data are represented as mean ± SD, *represents P < 0.05, **represents P < 0.01

Article Snippet: The primary antibodies, including alpha synuclein antibody (α-syn, 10842-1-AP, Proteintech, China), TH antibody (25859-1-AP, Proteintech, China), FTO antibody (27226-1-AP, Proteintech, China), and ATM antibody (27156-1-AP, Proteintech, China) were used.

Techniques: In Vivo, Confocal Microscopy, Quantitative RT-PCR, Injection, Western Blot, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson’s disease via the microglial NLRP3 inflammasome

doi: 10.1186/s12974-022-02494-y

Figure Lengend Snippet: Diurnal changes in microglia and the inflammatory-related genes in SN of MPTP-induced Mice. A Representative images of IBA1 staining in the SN of control and MPTP mouse with IBA1 size quantification ( B ). Scale bar, 50 μm. Mice were killed at ZT6 and ZT18. C Iba1 staining in the control and MPTP group with Iba1 number quantification ( D ). Scale bar, 200 μm. E–H RT-PCR analysis for microglial inflammatory genes from SN of Control and MPTP mice. n = 3–4 for each group. Data were presented as mean ± SEM. (* p < 0.05, ** p < 0.01, or **** p < 0.0001 by two-tailed t test)

Article Snippet: For immunohistochemical staining, the primary antibodies used were as follows: rabbit anti-TH (25859-1-AP, Proteintech), rabbit anti-IBA1 (NO. 019-19741, Wako), and mouse anti-GFAP (60190-1-Ig, Proteintech).

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson’s disease via the microglial NLRP3 inflammasome

doi: 10.1186/s12974-022-02494-y

Figure Lengend Snippet: SR9009 inhibits microgliosis and astrocytosis in the SN of MPTP-induced mice. The representative immunohistochemistry staining ( A ) and the statistical graph ( B ) of IBA1 in the SN. Scale bars, 250 μm for the top row and 50 μm for the bottom row. The representative immunohistochemistry staining ( C ) and the statistical graph ( D ) of GFAP in the SN. Scale bars, 500 μm for the top row and 100 μm for the bottom row. Representative western blot bands ( E ) and the statistical graph ( F , G ) of IBA1 and GFAP in the SN. The protein levels were normalized to β-actin. n = 3–4 for each group. Data were presented as mean ± SEM. (* p < 0.05, ** p < 0.01, or *** p < 0.001 by one-way ANOVA test)

Article Snippet: For immunohistochemical staining, the primary antibodies used were as follows: rabbit anti-TH (25859-1-AP, Proteintech), rabbit anti-IBA1 (NO. 019-19741, Wako), and mouse anti-GFAP (60190-1-Ig, Proteintech).

Techniques: Immunohistochemistry, Staining, Western Blot

Journal: Journal of Neuroinflammation

Article Title: The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson’s disease via the microglial NLRP3 inflammasome

doi: 10.1186/s12974-022-02494-y

Figure Lengend Snippet: SR9009 inhibits microgliosis and astrocytosis in striatum of MPTP-induced mice. A The representative immunohistochemistry staining of IBA1 in the striatum. Scale bars, 1 mm for the top row and 100 μm for the bottom row. B The representative immunohistochemistry staining of GFAP in the striatum. Scale bars, 1 mm for the top row and 100 μm for the bottom row. C , D The statistical graph of IBA1 and GFAP in the striatum. (* p < 0.05, ** p < 0.01, or *** p < 0.001 by One-way ANOVA test)

Article Snippet: For immunohistochemical staining, the primary antibodies used were as follows: rabbit anti-TH (25859-1-AP, Proteintech), rabbit anti-IBA1 (NO. 019-19741, Wako), and mouse anti-GFAP (60190-1-Ig, Proteintech).

Techniques: Immunohistochemistry, Staining

Journal: Journal of Neuroinflammation

Article Title: The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson’s disease via the microglial NLRP3 inflammasome

doi: 10.1186/s12974-022-02494-y

Figure Lengend Snippet: SR9009 reversed the phenotypic polarization of microglia in the SN of MPTP-induced mice. A Representative double-immunofluorescent staining of IBA1 (green) and iNOS (red) in the SN. Scale bar, 50 μm. D Representative double-immunofluorescent staining of IBA1 (green) and iNOS (Arg-1) in the SN. Scale bar, 50 μm. The real-time PCR results of iNOS ( B ), Arg-1 ( C ), TNF-α ( E ) and IL-6 ( F ) in the SN. n = 3–4 for each group. Data were presented as mean ± SEM. (* p < 0.05, ** p < 0.01, or *** p < 0.001 by One-way ANOVA test)

Article Snippet: For immunohistochemical staining, the primary antibodies used were as follows: rabbit anti-TH (25859-1-AP, Proteintech), rabbit anti-IBA1 (NO. 019-19741, Wako), and mouse anti-GFAP (60190-1-Ig, Proteintech).

Techniques: Staining, Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson’s disease via the microglial NLRP3 inflammasome

doi: 10.1186/s12974-022-02494-y

Figure Lengend Snippet: SR9009 suppresses NLRP3 inflammasome activation in the SN of MPTP-induced mice. The representative western blot bands ( A ) and the statistical graph ( B ) of p-NF-κB p65 and NF-κB p65 in the SN. The representative western blot bands ( E ) and the statistical graph ( C – D , F ) of NLRP3, ASC and cleaved-caspase-1 in the SN. The real-time PCR results of IL-1β ( G ) and IL-18 ( H ) in the SN. I Representative double-immunofluorescent staining of IBA1 (red) and NLRP3 (green) in the SN and the statistical graph ( J ) of IBA1 + NLRP3 + /IBA1 + cells. Scale bar, 50 μm. n = 3–4 for each group. The p-NF-κB p65 level was normalized to the total of NF-κB p65, and the rest protein levels were normalized to β-actin. Data were presented as mean ± SEM. (* p < 0.05, ** p < 0.01, or *** p < 0.001 by One-way ANOVA test)

Article Snippet: For immunohistochemical staining, the primary antibodies used were as follows: rabbit anti-TH (25859-1-AP, Proteintech), rabbit anti-IBA1 (NO. 019-19741, Wako), and mouse anti-GFAP (60190-1-Ig, Proteintech).

Techniques: Activation Assay, Western Blot, Real-time Polymerase Chain Reaction, Staining