Journal: Neural Regeneration Research

Article Title: Tranylcypromine upregulates Sestrin 2 expression to ameliorate NLRP3-related noise-induced hearing loss

doi: 10.4103/NRR.NRR-D-24-00130

Figure Lengend Snippet: TCP increases SESN2 levels in NE mice cochleae. (A) Representative images of the medial cochlear turn in the control, NE+saline, and NE+TCP groups ( n = 3 cochleae from three mice per group). Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for SESN2 (green), and nuclei (DAPI; blue). (B) Hair cells were visualized by immunofluorescent staining for phalloidin (red), and the tissues were also stained for NLRP3 (green), and nuclei (DAPI; blue) (n=3 cochleae from three mice per group). Scale bars: 20 μm in A and B. (C) Quantification of SESN2- and NLRP3-positive cells among the three groups. (D) SESN2 expression in mice cochleae ( n = 3 cochleae from three mice per group). All experiments were replicated thrice. Data are presented as mean ± standard of the mean. * P < 0.05, ** P < 0.01, **** P < 0.0001 (one-way analysis of variance with Bonferroni’s multiple comparison test). Ctrl: Control; DAPI: 4,6-diamidino-2-phenylindole; NE: noise exposure; ns: not significant; TCP: tranylcypromine.

Article Snippet: First, the membranes were incubated overnight at 4°C with the following primary antibodies: mouse monoclonal anti-SESN2 (1:200, sc-393195, Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit polyclonal anti-NLRP3 (1:400, NBP2-12446, Novus Biologicals, Centennial, CO, USA), rabbit polyclonal anti-LC3B (1:250, ab192890, Abcam, Cambridge, UK), rat monoclonal anti-LAMP1 (1:200, 14-1071-82, Invitrogen, Waltham, MA, USA), and mouse monoclonal 4-hydroxynonenal (4-HNE) (1:250, MA5-27570, Invitrogen).

Techniques: Control, Saline, Staining, Expressing, Comparison

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Reagents and instruments used in the study

Article Snippet: Rabbit polyclonal anti-HO-1 antibody , Proteintech Group , 10701-1-AP (RRID: AB_2923713).

Techniques: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Reagents and instruments used in the study

Article Snippet: Rabbit polyclonal anti-PSD95-specific antibody , Proteintech Group , 20665-1-AP (RRID: AB_2687961).

Techniques: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Treatment with NaHS inhibits the dysregulation of neuroplasticity induced by Quin. (A, B) Immunofluorescence images showing colocalization of PSD95 (red, CoraLite594 fluorescent dye) and NeuN (green, CoraLite488 fluorescent dye) in nine coronal sections from three mice per group. PSD95 expression was significantly upregulated in neuronal cells in the Quin + NaHS group compared with the Quin group, and this effect was reversed by the addition of ML385. Scale bar: 20 μm. (C) Western blot was used to measure PSD95, SYT1, and Syn expression levels 6 days after Quin striatal injection. Quantification of PSD95, SYT1, and Syn expression levels ( n = 3 per group). The values are reported as mean ± SD. *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). NeuN: Neuronal nuclei; PSD95: postsynaptic density protein 95; Quin: quinolinic acid; Syn: synaptophysin; SYT1: synaptotagmin 1.

Article Snippet: Rabbit polyclonal anti-PSD95-specific antibody , Proteintech Group , 20665-1-AP (RRID: AB_2687961).

Techniques: Immunofluorescence, Expressing, Western Blot, Injection

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Reagents and instruments used in the study

Article Snippet: Rabbit polyclonal anti-DAO antibody , Proteintech Group , 13273-1-AP (RRID: AB_ 2877932).

Techniques: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Reagents and instruments used in the study

Article Snippet: Rabbit polyclonal anti-NRF2 antibody , Proteintech Group , 16396-1-AP (RRID: AB_2782956).

Techniques: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: NaHS reduces oxidative stress by suppressing Nrf2. (A) Representative images of DCFHDA staining in PC12 cells treated with Quin for 24 hours. The Quin group exhibited greater DCFH-DA (green) fluorescence than the Sham group. The Quin + NaHS group exhibited a decrease in DCFHDA (green) fluorescence compared with the Quin group, while an increase was observed in the Quin + NaHS + ML385 group. Scale bar: 50 μm. (B) DHE staining of PC12 cells following 24-hour Quin stimulation. Stronger DHE (red) fluorescence was observed in the Quin group compared with the Sham group. DHE (red) fluorescence was decreased in the Quin + NaHS group and increased in the Quin + NaHS + ML385 group compared with the Quin group. Scale bar: 50 μm. (C) DCFH-DA fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (D) DHE fluorescence intensity in PC12 cells was quantified using a fluorescence microplate reader ( n = 5 per group). (E) DCFHDA fluorescence intensity in the striatum was quantified using a fluorescence microplate reader ( n = 5 per group). (F) Western blot detection of Nrf2 and HO-1 expression levels in PC12 cells in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). (G) Western blot detection of Nrf2 and HO-1 expression levels in the striatum of mice in each group. Quantification of Nrf2 and HO-1 expression levels ( n = 3 per group). The values are reported as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). DCFHDA: 2′,7′-Dichlorodihydrofluorescein diacetate; DHE: dihydroethidium; HO-1: heme oxygenase-1; Nrf2: nuclear factor erythroid 2-related factor 2; Quin: quinolinic acid.

Article Snippet: Rabbit polyclonal anti-NRF2 antibody , Proteintech Group , 16396-1-AP (RRID: AB_2782956).

Techniques: Staining, Fluorescence, Western Blot, Expressing

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: NaHS regulates mitochondrial function in Quin-stimulated PC12 cells by suppressing Nrf2. (A) Mitochondria stained with MitoTracker (red) in Quin-stimulated PC12 cells treated with NaHS and ML385. The Quin group exhibited severe mitochondrial fragmentation and a decrease in mitochondrial fluorescence intensity compared with the Sham group, whereas the addition of NaHS resulted in an increase in mitochondrial fluorescence intensity. However, this effect was reversed by the addition of ML385. Scale bar: 10 μm. (B) Quantification of mitochondrial fluorescence intensity ( n = 10 per group). (C) Representative micrographs of MMP in Quin-stimulated PC12 cells, as detected via JC-1. The transition from green to red fluorescence indicates mitochondrial polarization, which was restored to normal levels by NaHS treatment. In contrast, ML385 treatment induced a shift from green to red fluorescence, indicating substantial mitochondrial depolarization. Scale bar: 50 μm. (D) Quantification of mitochondrial MMP ( n = 15 per group). (E) Quantification of mitochondrial DNA (mtDNA, NADH5) versus nuclear DNA (GAPDH) as detected by real-time polymerase chain reaction in Quin-stimulated PC12 cells treated with NaHS and ML385. (F) Western blot detection of TOMM20 expression levels in PC12 cells. Quantification of TOMM20 expression ( n = 3 per group). The values are reported as mean ± SD. ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Bonferroni post hoc test). GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; MMP: mitochondrial membrane potential; Quin: quinolinic acid; TOMM20: translocase of outer mitochondrial membrane 20.

Article Snippet: Rabbit polyclonal anti-NRF2 antibody , Proteintech Group , 16396-1-AP (RRID: AB_2782956).

Techniques: Staining, Fluorescence, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Membrane

Journal: Neural Regeneration Research

Article Title: Hydrogen sulfide reduces oxidative stress in Huntington’s disease via Nrf2

doi: 10.4103/NRR.NRR-D-23-01051

Figure Lengend Snippet: Reagents and instruments used in the study

Article Snippet: Rabbit polyclonal anti-CBS antibody , Proteintech Group , 14787-1-AP (RRID: AB_2070970).

Techniques: CCK-8 Assay, Saline, TUNEL Assay, DNA Extraction, Modification, Marker, Fluorescence, Microscopy, Imaging