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. 2021 Jun 11;12:657873. doi: 10.3389/fendo.2021.657873

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Copyright © 2021 Alhaidan, Christesen, Lundberg, Balwi and Brusgaard

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Adcy7 knockout establishment. (A) Sanger sequencing wild-type RIN-m⁽ ^Adcy7 ^/ ^Adcy7 ⁾ (Top) compared to the mutant cell line (bottom) confirmed an insertion creating a frameshift. (B) Amino acid sequencing with a reference (top) shows indel creating a frameshift causing a premature termination codon located in 11 codons downstream. (C) Adcy7 mRNA levels established by quantitative RT-PCR indicating a very low expression level. The CT values were normalized to β-actin to yield ΔCT and 2^-ΔΔCT values presented in dots and bar chart, respectively. Each dot represents an individual sample, whereas the horizontal lines represent the mean and standard error of the mean. ΔCT values were then calculated and presented as 2^-ΔΔCT values (fold change) in a bar chart. Unpaired t-test with delta CT and SE ± values were used for statistic test.