antibodies against myo7a  (Covance)

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV-regulated Serpine2 overexpression promotes hair cell regeneration

doi: 10.1016/j.omtn.2024.102396

Figure Lengend Snippet: Serpine2 overexpression promoted cochlear progenitor differentiation into HCs in two- and three-dimensional cultures in vitro (A) Experimental design (two-dimensional differentiation assay). (B) Immunofluorescence staining of differentiated organoids inside and outside of the colony in control and AAV- Serpine2 overexpression groups. Myo7a (red) marks HCs. AAV-Green (green) marks cells transduced by virus. Scale bar, 50 μm. (C) The numbers of Myo7a + /Green + cells inside or outside the organoid per well in (B). (D) Experimental design (three-dimensional differentiation assay). (E) Immunofluorescence staining of differentiated organoids in the control and AAV- Serpine2 overexpression groups. Myo7a (red) marks HCs. AAV-Green (green) marks cells transduced by virus. Scale bar, 100 μm. (F–H) The number of Myo7a + organoids, the percentage of Myo7a + organoids, and the amount of Myo7a + cells in every well in (E).

Article Snippet: The samples were incubated with primary antibodies against Myo7a (Proteus Biosciences, #25–6790, 1:1000 dilution) and Sox2 (Santa, #SC-17320, 1:400 dilution) for 12 h, and then they were incubated with the corresponding secondary antibody.

Techniques: Over Expression, In Vitro, Differentiation Assay, Immunofluorescence, Staining, Control, Virus

Journal: Molecular Therapy. Nucleic Acids

Article Title: AAV-regulated Serpine2 overexpression promotes hair cell regeneration

doi: 10.1016/j.omtn.2024.102396

Figure Lengend Snippet: Serpine2 overexpression promoted Lgr5 + progenitor cell trans -differentiation into HCs in vivo (A) Experimental design. P1 Lgr5-EGFP CreER/+ /Rosa26-tdTomato loxp/+ mice were injected with tamoxifen intraperitoneally. AAV viruses were injected 12 h later through the RWM, and cochleae were collected at P7. (B) Immunofluorescence images of cochlear epithelia transduced by control and AAV- Serpine2 viruses. Myo7a + /tdTomato + HCs are marked by yellow arrows. Scale bar, 50 μm. (C) The number of Myo7a + /tdTomato + OHCs, IHCs, and HCs in every cochlea.

Article Snippet: The samples were incubated with primary antibodies against Myo7a (Proteus Biosciences, #25–6790, 1:1000 dilution) and Sox2 (Santa, #SC-17320, 1:400 dilution) for 12 h, and then they were incubated with the corresponding secondary antibody.

Techniques: Over Expression, In Vivo, Injection, Immunofluorescence, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: PGC-1 α -mediated imbalance of mitochondria-lipid droplet homeostasis in neomycin-induced ototoxicity and nephrotoxicity

doi: 10.1016/j.apsb.2024.05.024

Figure Lengend Snippet: The characteristics of neomycin distribution in the cochlea and kidney correlate with ototoxicity and nephrotoxicity. (A) Schematic diagram of 200 mg/kg neomycin administration for 7 days followed by 7 days drug withdrawal in mice. (B) Histological photomicrographs of kidney tissue sections stained with H&E, PAS. Scale bar = 50 μm. (C) Analysis of serum creatinine, Ngal and Kim- 1 mRNA levels ( n = 4–6). (D) Representative confocal images of hair cells. Scale bar = 20 μm. (E) Quantification of MYO7A + hair cells of three turns of cochleae ( n = 3). (F) ABR measurement of mice ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 D8 vs . control group. # P < 0.05, ## P < 0.01, ### P < 0.001 D14 vs . control group. (G) Representative confocal images depicting NTR accumulation in various organs of mice. Scale bar = 20 μm. (H) Determination of drug concentration in various tissues detected by HPLC–MS/MS ( n = 9). Con: mice treated with saline once daily for 7 days; D4: mice treated with 200 mg/kg neomycin once daily subcutaneously for 3 days; D8: mice treated with 200 mg/kg neomycin once daily subcutaneously for 7 days; D15: mice treated with 200 mg/kg neomycin once daily subcutaneously for 7 days followed by drug withdrawal for 7 days. (I) Representative confocal images of three turns of the cochlear explants. Scale bar = 20 μm. (J) Quantification of MYO7A + hair cells of the cochlear explants ( n = 3). (K) Quantification of fluorescence area of NTR in cochlear explants ( n = 3). Con, cochlear explants cultured with culture media for 18 h; Neo, cochlear explants treated with 0.5 mmol/L neomycin for 6 h followed by 12 h recovery. Ngal , neutrophil gelatinase-associated lipocalin; Kim-1 , kidney injury molecule 1. The results are presented as the means ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: The membranes were then incubated with primary antibodies against MYO7A (1:400, Proteus Biosciences Inc, Waltham, MA, USA) diluted in blocking solution at 4 °C overnight.

Techniques: Staining, Control, Concentration Assay, Tandem Mass Spectroscopy, Saline, Fluorescence, Cell Culture

Journal: Acta Pharmaceutica Sinica. B

Article Title: PGC-1 α -mediated imbalance of mitochondria-lipid droplet homeostasis in neomycin-induced ototoxicity and nephrotoxicity

doi: 10.1016/j.apsb.2024.05.024

Figure Lengend Snippet: Pgc-1α −/− aggravated neomycin-induced toxicity and lipid metabolism disorder in vivo . (A) Schematic diagram of 100 mg/kg neomycin treatment for 7 consecutive days in mice. (B) Analysis of serum creatinine ( n = 5), as well as relative RNA expression of renal tubular injury marker genes ( n = 7–8). (C) Histological photomicrographs of kidney tissue sections stained with H&E, PAS, Scale bar = 50 μm; NGAL immunostaining of kidney tissue sections, Scale bar = 50 μm; Electron micrographs of mitochondria in kidney tubule. Scale bar = 2 μm. (D) ABR measurement of mice ( n = 3). (E) Representative confocal images of hair cells and quantification of MYO7A + hair cells in three turns of cochleae ( n = 3). Scale bar = 20 μm. (F) Heatmaps and counts of significantly changed lipids ( P < 0.05) in inner ears and kidneys ( n = 4–6). (G) Volcano plot illustrating significantly altered TAGs estimated by LC–MS. (H) Relative abundance for TAGs and PLs containing double bonds by LC–MS. Experiment groups: WT + Neo, WT mice treated with 100 mg/kg neomycin once daily subcutaneously for 7 consecutive days; Pgc-1α −/− +Neo, Pgc-1α −/− mice treated with 100 mg/kg neomycin once daily subcutaneously for 7 consecutive days. Ngal , neutrophil gelatinase-associated lipocalin; Kim-1 , hepatitis A virus cellular receptor 1. The results are presented as the mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: The membranes were then incubated with primary antibodies against MYO7A (1:400, Proteus Biosciences Inc, Waltham, MA, USA) diluted in blocking solution at 4 °C overnight.

Techniques: In Vivo, RNA Expression, Marker, Staining, Immunostaining, Liquid Chromatography with Mass Spectroscopy, Virus