Journal: Scientific Reports

Article Title: Involvement of Cl − /HCO 3 − exchanger SLC26A3 and SLC26A6 in preimplantation embryo cleavage

doi: 10.1038/srep28402

Figure Lengend Snippet: ( A ) Immunofluorescent results showing the localization of SLC26A3 (upper panel, green) and SLC26A6 (lower panel, green) protein in different stages of human and mouse preimplantation embryos. Nuclei were counterstained with DAPI (blue). ( B–E ) Quantitative real-time PCR results showing the levels of SLC26A3 ( B , D ) and SLC26A6 ( C , E ) mRNA expression during human ( B,C ) and mouse ( D,E ) preimplantation embryo development. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001 (by one-way ANOVA, ~100 embryos in each experiment, n = 4).

Article Snippet: The siRNA negative control (siRNA NC, sc-37007, Santa Cruz Biotechnology, CA, USA), SLC26A3 siRNA (sc-45544, Santa Cruz Biotechnology) or SLC26A6 siRNA (sc-108024, Santa Cruz Biotechnology) was dissolved in RNase-free water.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Scientific Reports

Article Title: Involvement of Cl − /HCO 3 − exchanger SLC26A3 and SLC26A6 in preimplantation embryo cleavage

doi: 10.1038/srep28402

Figure Lengend Snippet: ( A–F ) Effects of inhibiting CFTR (CFTRinh172, 10 μM), SLC26A3 (niflumate, 20 μM) and SLC26A6 (DIDS, 20 μM) on preimplantation embryo cleavage in complete TALP ( A,B ) control: 32/38 embryos; CFTRinh172: 6/34 embryos; niflumate: 9/35 embryos; DIDS: 14/40 embryos), HCO 3 − -deficient TALP ( C,D ) control: 5/30 embryos; CFTRinh172: 7/31 embryos; niflumate: 4/32 embryos; DIDS: 6/31 embryos) and Cl − -deficient TALP ( E,F , control: 6/32 embryos; CFTRinh172: 4/31 embryos; niflumate: 5/30 embryos; DIDS: 6/33 embryos). Scale bar: 100 μm. Summary of the results are shown on the right panel ( B , D , F ). ns indicates P > 0.05, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001 (by one-way ANOVA, n = 4). ( G–I ) Quantitative real-time PCR showing the expression of miRNA-125b ( G ), p53 ( H ) and p21 ( I ) after CFTRinh172, niflumate or DIDS treatment. * indicates P < 0.05, ** indicates P < 0.01 (by one-way ANOVA, ~100 embryos in each experiment, n = 4).

Article Snippet: The siRNA negative control (siRNA NC, sc-37007, Santa Cruz Biotechnology, CA, USA), SLC26A3 siRNA (sc-45544, Santa Cruz Biotechnology) or SLC26A6 siRNA (sc-108024, Santa Cruz Biotechnology) was dissolved in RNase-free water.

Techniques: Control, Real-time Polymerase Chain Reaction, Expressing

Journal: Scientific Reports

Article Title: Involvement of Cl − /HCO 3 − exchanger SLC26A3 and SLC26A6 in preimplantation embryo cleavage

doi: 10.1038/srep28402

Figure Lengend Snippet: (A,B ) Quantitative real-time PCR result showing the expression of SLC26A3 ( A ) and SLC26A6 ( B ) mRNA in embryos after control siRNA (siRNA NC), SLC26A3 siRNA, SLC26A6 siRNA or both SLC26A3 and SLC26A6 siRNA injection at the 2-cell stage. *** indicates P < 0.001 (by one-way ANOVA, n = 4). ( C ) Four-cell embryo formation after 12 hours of embryo culture following siRNA NC, SLC26A3 siRNA, SLC26A6 siRNA or both SLC26A3 and SLC26A6 siRNA injection at 2-cell stage (siRNA NC: 32/45 embryos; SLC26A3 siRNA: 11/44 embryos; SLC26A6 siRNA: 21/40 embryos; SLC26A3 + A6 siRNA: 5/42 embryos). Scale bar: 100 μm. ( D ) Summary of the results from C. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001 (by one-way ANOVA, n = 4). ( E–G ) Quantitative real-time PCR showing the expression of miRNA-125b ( E ), p53 ( F ) and p21 ( G ) after siRNA NC, SLC26A6 siRNA, SLC26A3 siRNA or both SLC26A3 and SLC26A6 siRNA injection. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001 (by one-way ANOVA, ~100 embryos in each experiment, n = 4).

Article Snippet: The siRNA negative control (siRNA NC, sc-37007, Santa Cruz Biotechnology, CA, USA), SLC26A3 siRNA (sc-45544, Santa Cruz Biotechnology) or SLC26A6 siRNA (sc-108024, Santa Cruz Biotechnology) was dissolved in RNase-free water.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Control, Injection, Embryo Culture

Journal: Scientific Reports

Article Title: Involvement of Cl − /HCO 3 − exchanger SLC26A3 and SLC26A6 in preimplantation embryo cleavage

doi: 10.1038/srep28402

Figure Lengend Snippet: Working model for the regulation of early embryo cleavage by SLC26A3 and SLC26A6 in CFTR/HCO 3 − -dependent activation of miR-125b. The HCO 3 − influx is mediated by SLC26A3 and SLC26A6 with an exchange of 2Cl − / HCO 3 − . Apart from its reported role in conducting HCO 3 − directly, CFTR act as a Cl − channel to provide a recycling pathway for Cl − that is required for SLC26A3 and SLC26A6 function. The sites of action for inhibitors CFTRinh172, niflumate, and DIDS, as well as the intracellular HCO 3 − -dependent events, are also shown. HCO 3 − influx mediated by the cooperative action of SLC26A3, SLC26A6 and CFTR activates soluble adenylyl cyclase (sAC) which increase the level of cAMP. This in turn activates PKA/NFkB signaling cascade which increases the expression of miR125b. Expression of miR125b is required for embryo cleavage through suppressing the expression of p53 and p21. Vm, membrane potential; [pH]i, intracellular pH.

Article Snippet: The siRNA negative control (siRNA NC, sc-37007, Santa Cruz Biotechnology, CA, USA), SLC26A3 siRNA (sc-45544, Santa Cruz Biotechnology) or SLC26A6 siRNA (sc-108024, Santa Cruz Biotechnology) was dissolved in RNase-free water.

Techniques: Activation Assay, Expressing, Membrane