Journal: BioMed Research International

Article Title: Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells

doi: 10.1155/2023/4191999

Figure Lengend Snippet: Characteristics of HEI-OC1 cells and the expression of Kir4.1. Cell growth curve and Kir4.1 expression levels were observed to determine the experimental condition in HEI-OC1 cells. (a) Cell growth curves were obtained at 33°C and 39°C for 14 days in HEI-OC1 cells. (b) The change in the expression level of Kir4.1 was confirmed by qRT-PCR. (c) Expression of Kir4.1 at 33°C and 39°C was confirmed at the protein level by western blot. (d, e) Immunofluorescence assay was performed to confirm the changes in the expression level of Kir4.1 on day 3 and day 7 of cells at 33°C and 39°C, respectively. Expression of Kir4.1 was enhanced at 39°C compared to 33°C. High magnification microscopy revealed that the expression was mainly at the cell membrane surface.

Article Snippet: After 24 hours, Kir4.1 siRNA (sc-42625, Santa Cruz Biotechnology, Dallas, TX, USA) was added (40 pmol).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Microscopy, Membrane

Journal: BioMed Research International

Article Title: Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells

doi: 10.1155/2023/4191999

Figure Lengend Snippet: Association of potassium channels Kir4.1, NKCC1, and KCNQ4 in HEI-OC1 cells. Western blot and qRT-PCR results to determine the association of Kir4.1, NKCC1, and KCNQ4 in HEI-OC1 cells. (a) Treatment of HEI-OC1 cells with either kanamycin or furosemide increased the expression of Kir4.1. (b) When furosemide was treated in HEI-OC1 cells, NKCC1 was blocked, and kanamycin was not affected. (c) When kanamycin was treated in HEI-OC1 cells, KCNQ4 was blocked, and there was no specific effect upon furosemide treatment. (d) Changes in expression levels of potassium channels in ototoxic drug-treated HEI-OC1 cells confirmed by western blotting.

Article Snippet: After 24 hours, Kir4.1 siRNA (sc-42625, Santa Cruz Biotechnology, Dallas, TX, USA) was added (40 pmol).

Techniques: Western Blot, Quantitative RT-PCR, Expressing

Journal: BioMed Research International

Article Title: Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells

doi: 10.1155/2023/4191999

Figure Lengend Snippet: Changes in HEI-OC1 cells upon Kir4.1 channel inhibition. (a) qRT-PCR results of HEI-OC1 cells cultured 39°C for 3 days following Kir4.1 inhibition. (b) Expression of Kir4.1 with ototoxic drug treatment in HEI-OC1 cells cultured at 39°C for 3 days following siRNA transfection. (c) CCK assay of cell viability following transfection with Kir4.1 siRNA. (d) CCK assay results of Kir4.1 channel inhibition HEI-OC1 cells after 72 h of treatment with ototoxic drugs.

Article Snippet: After 24 hours, Kir4.1 siRNA (sc-42625, Santa Cruz Biotechnology, Dallas, TX, USA) was added (40 pmol).

Techniques: Inhibition, Quantitative RT-PCR, Cell Culture, Expressing, Transfection

Journal: BioMed Research International

Article Title: Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells

doi: 10.1155/2023/4191999

Figure Lengend Snippet: Changes in Kir4.1 expression in control and ototoxic mice. (a–d) Kir4.1 expression in control mice. (a) Expression of Kir4.1 in stria vascularis (StV) and outer hair cells (OHC) of control mice. (b) Pattern of staining with 4′,6-diamidino-2-phenylindole (DAPI). (c) Merged image. (d) High expression of Kir4.1 in OHC. (e–h) Kir4.1 expression in ototoxic mice. (e) Expression of Kir4.1 in StV and OHC of ototoxic mice. (f) Pattern of DAPI staining. (g) Merged image. (h) Organ of Corti displayed decreased intensity expression of Kir4.1 in OHC.

Article Snippet: After 24 hours, Kir4.1 siRNA (sc-42625, Santa Cruz Biotechnology, Dallas, TX, USA) was added (40 pmol).

Techniques: Expressing, Staining

Journal: BioMed Research International

Article Title: Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells

doi: 10.1155/2023/4191999

Figure Lengend Snippet: Changes in expression of Kir4.1 ex vivo . Kir4.1 expression in outer hair cells and inner hair cells (OHC) in cochlear explants of control and ototoxic mice. (a–c) Explant of the control mouse. (a) Kir4.1 is strongly expressed in OHC. (b) Upon DAPI staining, cell loss was observed. (d–f) Explant of ototoxic mouse. (d) The expression of Kir4.1 in OHC was decreased compared to the control. (e) Upon DAPI staining, cell loss was observed in OHC.

Article Snippet: After 24 hours, Kir4.1 siRNA (sc-42625, Santa Cruz Biotechnology, Dallas, TX, USA) was added (40 pmol).

Techniques: Expressing, Ex Vivo, Staining

Journal: BioMed Research International

Article Title: Role of Kir4.1 Channels in Aminoglycoside-Induced Ototoxicity of Hair Cells

doi: 10.1155/2023/4191999

Figure Lengend Snippet: Correlation of Kir4.1, NKCC1, and KCNQ4 in HEI-OC1 cells. Correlation between the main potassium channels Kir4.1, NKCC1, and KCNQ4. When NKCC1 is blocked, the Kir4.1 channel has compensatory action and allows increased influx of potassium into cells. Kir4.1 is normally an inwardly rectifying K + potassium channel. However, when the KCNQ4 channel is blocked, Kir4.1 channel allows efflux of K + instead of through the KCNQ4 channel.

Article Snippet: After 24 hours, Kir4.1 siRNA (sc-42625, Santa Cruz Biotechnology, Dallas, TX, USA) was added (40 pmol).

Techniques: