Journal: Bioengineered

Article Title: Small activating RNA activation of ATOH1 promotes regeneration of human inner ear hair cells

doi: 10.1080/21655979.2022.2045835

Figure Lengend Snippet: Screening of ATOH1 small activating RNAs in 293 T cells and mesenchymal stem cells . Cells were transfected with the indicated small activating RNAs using RNAiMAX at the indicated concentrations for 72 h. Mock samples were transfected in the absence of duplex RNA. Plasmid DNA was transfected at 1 µg/µl using Lipofectamine 3000. The transfected cells were subjected to RT–qPCR after RNA isolation, an RT reaction to generate cDNA and a Western blotting assay after protein isolation. A and B. Relative mRNA expression levels of the ATOH1 gene in 293 T cells (a) and mesenchymal stem cells (b) 72 h after transfection. C and D. ATOH1 protein levels assessed by Western blotting in 293 T cells (c) and mesenchymal stem cells (d) at 72 h after transfection. E and F. Relative ATOH1 mRNA expression levels in response to different small activating RNA concentrations.

Article Snippet: The human embryonic kidney cell line 293 T (ATCC® CRL) and human adipose-derived mesenchymal stem cells (ScienCell) were used as the cell lines to screen the small activating RNAs.

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Isolation, Western Blot, Expressing

Journal: Bioengineered

Article Title: Small activating RNA activation of ATOH1 promotes regeneration of human inner ear hair cells

doi: 10.1080/21655979.2022.2045835

Figure Lengend Snippet: Induction of hair cell progenitor cells . Mesenchymal stem cells were cultured in induction medium containing EGF, IGF-1 and bFGF for 4 weeks, and hair cell progenitor cell marker genes were detected. A . Mesenchymal stem cells were cultured in hair cell progenitor cell induction medium as described in the Materials and Methods. Total cellular RNA was isolated from the treated cells and reverse transcribed into cDNA, which was amplified by semiquantitative RT–PCR. The GAPDH gene was also amplified as a control for RNA loading. B. Treated cells were fixed and stained with an antibody against PAX8, and the nuclei were counterstained. Red arrows denote positively stained cells.

Article Snippet: The human embryonic kidney cell line 293 T (ATCC® CRL) and human adipose-derived mesenchymal stem cells (ScienCell) were used as the cell lines to screen the small activating RNAs.

Techniques: Cell Culture, Marker, Isolation, Reverse Transcription, Amplification, Reverse Transcription Polymerase Chain Reaction, Control, Staining