goat anti-slc12a2  (Santa Cruz Biotechnology)

Journal: Frontiers in Molecular Neuroscience

Article Title: Single-Cell RNA-Seq of Cisplatin-Treated Adult Stria Vascularis Identifies Cell Type-Specific Regulatory Networks and Novel Therapeutic Gene Targets

doi: 10.3389/fnmol.2021.718241

Figure Lengend Snippet: Expression of key EP-related cell-type specific proteins is reduced in the SV after cisplatin treatment. (A) Twenty four hours after cisplatin injection, SLC12A2 immunofluorescence intensity was significantly reduced in the apical and basal turns of the cochlea (two-way ANOVA, p < 0.0001; Sidak’s multiple comparisons test, q = 0.0068 and q = 0.0022, respectively). (B,C) Representative SLC12A2 immunostaining of SV cross sections of control and cisplatin-treated mice. (D) Immunofluorescence staining intensity of KCNJ10 was unchanged between control and cisplatin-treated mice (two-way ANOVA, p = 0.1784). (E,F) Representative KCNJ10 immunostaining of SV cross sections of control and cisplatin-treated mice. White arrows point to representative intermediate cell nuclei. (G) Immunofluorescence staining intensity of CLDN11 was reduced in the basal turn of cisplatin-treated compared to control mice (two-way ANOVA, p = 0.0002; Sidak’s multiple comparisons test q = 0.0418). (H,I) Representative CLDN11 immunostaining of SV cross sections of control and cisplatin-treated mice. Scale bars: 20 μm. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Primary antibodies used included rabbit anti-KCNJ10 ( RRID:AB_2040120 , Alomone Labs, APC-035, polyclonal, dilution 1:200), rabbit anti-CLDN11 ( RRID:AB_2533259 , Life Technologies, 364500, polyclonal, dilution 1:200), goat anti-SLC12A2 ( RRID:AB_2188633 , Santa Cruz Biotech, sc-21545, polyclonal, dilution 1:200), and Phalloidin AlexaFluor 647 ( RRID: AB_2620155 , Invitrogen, A22287, dilution 1:250).

Techniques: Expressing, Injection, Immunofluorescence, Immunostaining, Staining

Journal: Frontiers in Molecular Neuroscience

Article Title: Single-Cell RNA-Seq of Cisplatin-Treated Adult Stria Vascularis Identifies Cell Type-Specific Regulatory Networks and Novel Therapeutic Gene Targets

doi: 10.3389/fnmol.2021.718241

Figure Lengend Snippet: ddPCR of dissected SV demonstrates a reduction in several EP-related genes after cisplatin treatment. In whole dissected SV, we observed a significant reduction in selected EP-related genes specific to marginal cells: Kcne1 ( q = 0.0020), Kcnq1 ( q = 0.0161), Atp1b2 ( q = 0.0020), and Slc12a2 ( q = 0.0020) (unpaired multiple t -test, Benjamini–Hochberg correction). A significant reduction in two EP-related genes specific to intermediate cells: Met (uncorrected p = 0.0064) and Kcnj13 (uncorrected p = 0.0117) was seen between control and cisplatin-treated mice (unpaired multiple t -test, Benjamini–Hochberg correction). Expression of other known intermediate cell-specific genes including Kcnj10 ( p = 0.1746) and Ednrb ( p = 0.5309) were not reduced in cisplatin-treated compared to control. Expression of EP-related genes in basal cells, including Gjb2 ( q = 0.0207) was significantly reduced, while Gjb6 ( q = 0.1086), and Cldn11 ( q = 0.4012), were not decreased in cisplatin-treated compared to control mice (unpaired multiple t -test, Benjamini–Hochberg correction). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Primary antibodies used included rabbit anti-KCNJ10 ( RRID:AB_2040120 , Alomone Labs, APC-035, polyclonal, dilution 1:200), rabbit anti-CLDN11 ( RRID:AB_2533259 , Life Technologies, 364500, polyclonal, dilution 1:200), goat anti-SLC12A2 ( RRID:AB_2188633 , Santa Cruz Biotech, sc-21545, polyclonal, dilution 1:200), and Phalloidin AlexaFluor 647 ( RRID: AB_2620155 , Invitrogen, A22287, dilution 1:250).

Techniques: Expressing

Journal: Genes

Article Title: Dynamic Spatiotemporal Expression Changes in Connexins of the Developing Primate’s Cochlea

doi: 10.3390/genes12071082

Figure Lengend Snippet: Comparison of the expression patterns of CX26 and CX30 at E115 and P0 in lateral wall fibrocytes. ( A – C ) At E115, both CX26 and CX30 expressions were detected in the upper half of the spiral ligament fibrocytes. At this stage, a relatively high expression of CX30 was observed. On the modiolus side (stria vascularis side), CX30 expression was predominantly observed, and, in several cells, only CX30 expression was detected (arrowhead in ( C )). CX26 expression was observed more laterally, and, on the lateral side of fibrocytes, only CX26 expression was detected (asterisk in ( C )). In the central region, overlapping expression of CX26 and CX30 was observed (arrow in ( C )). ( D – E ) At P0, both CX26 and CX30 were observed in the lateral wall fibrocytes and lateral side membrane of the basal cells of stria vascularis. In contrast to E115, the expression of CX30 was observed more broadly. On the lateral side of fibrocytes (type III fibrocytes), only CX30 expression was observed (asterisk in ( E )). SLC12A2 (NKCC1) was used to label stria vascularis and lateral wall fibrocytes. Nuclei were counterstained with Hoechst stain (blue). Scale bar: 100 µm in ( A , D ), 50 µm in ( B , E ), 20 µm in ( C ). StV: stria vascularis, OC: organ of Corti. ( A – E ): basal turn.

Article Snippet: Invitrogen, Carlsbad, CA, USA, #13-8100), rabbit anti-connexin 30 antibody (Z-PP9) (#71-2200, 1:500, Invitrogen, Carlsbad, CA, USA), mouse anti-myosin7a antibody (#138-1-s, 1:30, DSHB, Iowa City, IA, USA), rabbit anti-myosin7a (#25-6790, 1:200, Proteus Biosciences, Ramona, CA, USA), goat anti-SOX2 antibody (AF2018, 1:200; R&D Systems, Minneapolis, MN, USA), and goat anti-NKCC1 (SLC12A2) antibody (sc21545, 1:300; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Comparison, Expressing, Membrane, Staining