Journal: iScience

Article Title: Heat promotes melanogenesis by increasing the paracrine effects in keratinocytes via the TRPV3/Ca 2+ /Hh signaling pathway

doi: 10.1016/j.isci.2023.106749

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Article Snippet: Rabbit monoclonal anti-DCT , ZENBIO , Cat#821374; SwissProtID:P40126.

Techniques: Recombinant, SYBR Green Assay, Software

Journal: PLOS ONE

Article Title: Round-window delivery of lithium chloride regenerates cochlear synapses damaged by noise-induced excitotoxic trauma via inhibition of the NMDA receptor in the rat

doi: 10.1371/journal.pone.0284626

Figure Lengend Snippet: A, B Cochlear tissue immunostained with specific antibodies for hair cells (myosin 7a, red) and ribbon synapses (CtBP2, green) in the 16 kHz region one week ( A ) and two weeks ( B ) after noise exposure. The arrow indicates the position of a pre-synapse. C, D Quantification of the number of ribbon synapses at the relevant frequencies of 8 kHz, 12 kHz, 16 kHz, and 32 kHz. Median plot of pre-synaptic count was compared among the experimental groups (noise + vehicle, noise + 1 mM LiCl, and noise + 2 mM LiCl). Error bars show 95% confidence intervals. Statistical significance is indicated by asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001 based on the Kruskal Wallis test. n = number of animals.

Article Snippet: For immunostaining, the cochlear pieces were blocked with 5% normal goat serum (NGS) in PBS and 0.3% Triton X-100 (TX) at room temperature for 1 hour followed by overnight incubation at 4°C with the following primary antibodies diluted in 1% NGS with 0.3% TX: 1) mouse (IgG1) anti-CtBP2 (C-terminal binding protein) at 1:500 (BD Transduction Labs Cat# 612044, RRID: AB_399431) to quantify pre-synaptic ribbons and 2) rabbit anti-myosin 7a at 1:200 (Proteus Biosciences Cat# 25–6790, RRID: AB_10015251) to delineate hair cells.

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Journal: bioRxiv

Article Title: ERBB2 is a Key Mediator in Hearing Restoration in Noise-Deafened Young Adult Mice

doi: 10.1101/838649

Figure Lengend Snippet: A-C. Representative 200x confocal images of 12 kHz IHCs from an age-matched, control mouse without noise exposure (A), a control mouse at 90 DPN (B), and a CA-ERBB2 mouse at 90 DPN (C). Immunolabeling for CTBP2 (red), GRIA2 (green) reveal paired synaptic structures, and immunolabeling for MYO7 (blue) reveals IHC morphology. Size bar: 5 microns. D-F. Similar 200x confocal images of IHCs from the 24 kHz region, also from an age-matched, control mouse without noise exposure (D), a control mouse at 90 DPN (E), and a CA-ERBB2 mouse at 90 DPN (F). Arrows indicate IHCs with gaps in their MYO7 staining (E, F). G-I. Amira 3-dimensional reconstructions from the same stacks shown in (A-C). Strong labeling for GRIA2 is coded green, while weak labeling is in blue. CTBP2 (red) and MYO7 (white) are also shown. J-L. Amira 3-dimensional reconstructions from the same stacks shown in (D-F). IHCs were rotated to be viewed from the same approximate angle.

Article Snippet: The following primary antibodies were used: mouse anti-CTBP2 (aka C-Terminal Binding Protein 2; 1:200; BD Transduction Laboratories; RRID:AB_399431), chicken anti-GFP (1:500; Abcam; RRID:AB_300798), rabbit anti-GLAST (aka EAAT1; 1:100; Abcam; RRID:AB_304334), mouse anti-GRIA2 (aka GluR2/GluA2; 1:2000; Millipore; RRID:AB_2113875), mouse anti-MAP2 (aka Microtubule-Associated Protein 2; 1:250; R&D Systems, Cat #: MAB8304, RRID:AB_2814693), rabbit anti-Myosin7 (MYO7) (1:200; Proteus; RRID:AB_10013626), mouse anti-Myosin7 (1:100; Developmental Studies Hybridoma Bank, RRID:AB_2282417), and goat anti-Oncomodulin (OCM) antibody (1:1000; Santa Cruz; RRID:AB_2267583).

Techniques: Immunolabeling, Staining, Labeling

Journal: bioRxiv

Article Title: ERBB2 is a Key Mediator in Hearing Restoration in Noise-Deafened Young Adult Mice

doi: 10.1101/838649

Figure Lengend Snippet: A. For 12 kHz IHCs, CTBP2+ ribbons were paired with GRIA2+ post-synaptic densities in Amira 3D reconstructions as described in the text. Data from age-matched control mice without noise exposure (“no noise”) are compared to data from control mice at 90 DPN (“control”) and data from CA-ERBB2 mice at 90 DPN (“CA-ERBB2”). Approximately 240 ribbons were identified for each condition, from 6-8 IHCs per biological replicate (3 replicates each). The size distributions of paired ribbons from each condition were plotted in violin graphs. Means are represented by a red dot and standard deviations by a red bar. Shapiro-Wilk normality tests revealed the data distribution to be non-normal. The size distribution of ribbons in control mice 90 DPN is not significantly different from similar mice without noise exposure (p=0.138, Kruskal-Wallis rank sum test with multiple comparison adjustment), whereas synapses in CA-ERBB2 mice 90 DPN differ from both control mice without noise exposure and control mice at 90 DPN (p=6.6 x10 -16 , 2.1 x10 -14 , respectively, Kruskal-Wallis rank sum test with multiple comparison adjustment). B. Synaptic size distributions for 24 kHz IHCs were obtained, similarly to those at 12 kHz, and plotted in violin graphs, with means represented by a red dot and standard deviations by a red bar. Data from age-matched control mice without noise exposure (“no noise”) are compared to data from control mice at 90 DPN (“control”) and data from CA-ERBB2 mice at 90 DPN (“CA-ERBB2”). Each distribution is significantly different from each other: “no noise” vs. “control”, p=3.8 x10 -6 , “no noise” vs. “CA-ERBB2”, p=6.6 x10 -16 , and “control” vs. “CA-ERBB2”, 5.2 x10 -13 . All tests, Kruskal-Wallis rank sum test with multiple comparison adjustment, 2-3 biological replicates per condition with 6-8 IHCs per replicate.

Article Snippet: The following primary antibodies were used: mouse anti-CTBP2 (aka C-Terminal Binding Protein 2; 1:200; BD Transduction Laboratories; RRID:AB_399431), chicken anti-GFP (1:500; Abcam; RRID:AB_300798), rabbit anti-GLAST (aka EAAT1; 1:100; Abcam; RRID:AB_304334), mouse anti-GRIA2 (aka GluR2/GluA2; 1:2000; Millipore; RRID:AB_2113875), mouse anti-MAP2 (aka Microtubule-Associated Protein 2; 1:250; R&D Systems, Cat #: MAB8304, RRID:AB_2814693), rabbit anti-Myosin7 (MYO7) (1:200; Proteus; RRID:AB_10013626), mouse anti-Myosin7 (1:100; Developmental Studies Hybridoma Bank, RRID:AB_2282417), and goat anti-Oncomodulin (OCM) antibody (1:1000; Santa Cruz; RRID:AB_2267583).

Techniques: