mouse igg1 anti myo7a  (Developmental Studies Hybridoma Bank)

Journal: iScience

Article Title: mTORC2 regulates auditory hair cell structure and function

doi: 10.1016/j.isci.2023.107687

Figure Lengend Snippet: KO confirmation of HC-RicKO mice (A) Hemizygous Myo15 - Cre +/− mice were crossed with homozygous Rictor fl/fl mice harboring LoxP sites upstream and downstream of exon 4 and 5, respectively. Forward (F) and reverse primers (R) were used to detect Cre-mediated recombination, resulting in a 280 bp PCR product after Cre-mediated recombination of the Rictor allele. (B) PCR using genomic DNA from dissected organ of Corti (OC), spiral ganglion (SG) or vestibular organ (vest; macular and cristae ampullaris organs pooled) tissue, separate for left (L) and right (R) inner ears. Genomic DNA from outer ear skin of both genotypes was used as negative control of Cre-mediated recombination. Genomic DNA from heart samples of tamoxifen-induced cardiomyocyte-specific Rictor KO mice was used as positive control (pos ctrl). No template control (NTC) contains all PCR reaction components including primers but no DNA. Successful Cre-mediated recombination of the Rictor allele was only found in OCs and vestibular organs of HC-RicKO mice. (C) Representative images (maximum intensity projections) of the medial cochlear turn from 4-week-old mice stained with an antibody against Akt-pSer473. Myosin7a antibody, phalloidin and nuclear DAPI staining visualize the hair cells. Scale bar for all figures = 20 μm. (D) Quantification of mean fluorescence intensity (MFI) in arbitrary units (a.u.) of the Akt-pSer473 signal intensity in the medial cochlear turn of 4-week-old mice. n = 3 mice per genotype. Results are presented as means ± SDs. Student’s t test, ∗p < 0.05.

Article Snippet: Mouse monoclonal anti-myosin7a IgG1 , Developmental Studies Hybridoma Bank , Cat#138-1s; RRID: AB_2282417.

Techniques: Negative Control, Positive Control, Staining, Fluorescence

Journal: iScience

Article Title: mTORC2 regulates auditory hair cell structure and function

doi: 10.1016/j.isci.2023.107687

Figure Lengend Snippet: Hearing loss precedes hair cell loss in HC-RicKO mice (A) Cochleograms showing inner hair cell (left graphs) and outer hair cell (right graphs) loss in 5 percent (%) distances from the Apex at indicated timepoints. Place-frequency map calculated with formula d = (LOG10((f+6.664)/9.8)/LOG10(10))/0.0092 (where d is the distance from the Apex in % and f the frequency in kHz). , , n = 3–4 mice (2 weeks), 4 mice (4 weeks), 3 mice (8 weeks), and 3–4 mice (12 weeks) per genotype. Results are presented as means ± SDs. Student’s t test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. (B) Representative images (maximum intensity projections) of the medial cochlear turn from 8-week-old mice of both genotypes. Hair cells are visualized with phalloidin, a Myosin7a antibody and nuclear DAPI staining. Scale bar for all figures = 20 μm.

Article Snippet: Mouse monoclonal anti-myosin7a IgG1 , Developmental Studies Hybridoma Bank , Cat#138-1s; RRID: AB_2282417.

Techniques: Staining

Journal: iScience

Article Title: mTORC2 regulates auditory hair cell structure and function

doi: 10.1016/j.isci.2023.107687

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-myosin7a IgG1 , Developmental Studies Hybridoma Bank , Cat#138-1s; RRID: AB_2282417.

Techniques: Recombinant, Saline, Electron Microscopy, Software, Imaging

Journal: eLife

Article Title: A gradient of Wnt activity positions the neurosensory domains of the inner ear

doi: 10.7554/eLife.59540

Figure Lengend Snippet:

Article Snippet: Antibody , Mouse monoclonal IgG1 anti-Myo7a (RRID: AB_2282417 ) , Developmental Studies Hybridoma Bank , Clone 138–1 , IF (1:500).

Techniques: Software, Recombinant, Plasmid Preparation, Clone Assay, Control, Cloning, Isolation, Concentration Assay, Sequencing

Journal: eLife

Article Title: A gradient of Wnt activity positions the neurosensory domains of the inner ear

doi: 10.7554/eLife.59540

Figure Lengend Snippet: ( a–a” ) Whole-mount (tiled maximum projection) views of an E7 chicken inner ear electroporated at E2 with a control vector (T2-mEGFP) and immunostained for Sox2 ( a’ ) and two hair cell markers, Myo7a and HCA (‘HC’ in all panels). ( a” ) All sensory organs are properly formed: posterior (pc), anterior (ac) and lateral (lc) cristae, saccule (sc), utricle (ut), basilar papilla (bp), and lagena ( l ). ( b–c” ) An inner ear transfected with T2-βcat-GOF. Note the absence of EGFP expression and severe defects in overall morphology of the vestibular system and basilar papilla; the remaining sensory patches are small and abnormally shaped ( b’ ). ( c–c” ) Higher magnification of the vestibular Sox2-positive patches containing Myo7a and HCA-expressing hair cells. ( d–f” ) An inner ear transfected with T2-βcat-LOF. ( d–d’’ ) Whole-mount (tiled maximum projection) views demonstrating the presence of numerous ectopic sensory patches with hair cells, and severe defects in inner ear morphology. ( e-e’ ) Higher magnification of the dorsal region, where transfected cells form ectopic sensory patches positive for Sox2 ( e’ ) and populated with Myo7a and HCA-expressing hair cells (arrowheads). ( f–f” ) In contrast, in ventral domains, EGFP-positive patches are devoid of Sox2 and hair cell markers expression. The only remaining Sox2-expressing patches are not transfected (arrows).

Article Snippet: The following antibodies were used: rabbit anti-Jagged 1 (Santa-Cruz Biotechnology, Dallas, TX; sc-8303; 1:200), rabbit anti-Sox2 (Abcam, UK; 97959, 1:500), mouse IgG1 monoclonal anti-Sox2 (BD Biosciences, San Jose, CA; 561469, 1:500), mouse IgG1 anti-Islet1 (Developmental Studies Hybridoma Bank, Iowa City, IA; Clone 39.3F7, 1:250), mouse IgG1 anti-HA-tag (Babco Inc, Richmond, CA; MMS-101R, 1:500), mouse IgG1 anti-Myo7a (Developmental Studies Hybridoma Bank, 1:500), and mouse IgG1 anti-HCA (a kind gift of Guy Richardson, 1:1000).

Techniques: Plasmid Preparation, Transfection, Expressing