ZNF70, a novel ILDR2-interacting protein, contributes to the regulation of HES1 gene expression

Watanabe, Kazuhisa; Nakayama, Kazuhiro; Ohta, Satoshi; Tago, Kenji; Boonvisut, Supichaya; Millings, Elizabeth J.; Fischer, Stuart G.; LeDuc, Charles A.; Leibel, Rudolph L.; Iwamoto, Sadahiko

doi:10.1016/j.bbrc.2016.06.124

Cited by 15 publications

(15 citation statements)

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“…Total RNA was isolated using acid guanidinium thiocyanate–phenol reagent . Complementary deoxyribonucleic acid synthesis was carried out using the Verso cDNA Kit (Thermo Scientific, Waltham, MA, USA) with random hexamer primers.…”

Section: Methodsmentioning

confidence: 99%

“…Complementary deoxyribonucleic acid synthesis was carried out using the Verso cDNA Kit (Thermo Scientific, Waltham, MA, USA) with random hexamer primers. Quantitative PCR (qPCR) assays were carried out using the ViiA7 Real‐Time PCR System and KAPA SYBER FAST ROX Low qPCR kit (Kapa Biosystems, Wilmington, MA, USA) . Relative gene expression levels were quantified by qPCR followed by normalization to the internal control gene 36B4 .…”

Section: Methodsmentioning

confidence: 99%

“…Total RNA was isolated using acid guanidinium thiocyanatephenol reagent. 12 Complementary deoxyribonucleic acid synthesis was carried out using the Verso cDNA Kit (Thermo Scientific, Waltham, MA, USA) with random hexamer primers. Quantitative PCR (qPCR) assays were carried out using the ViiA7 Real-Time PCR System and KAPA SYBER FAST ROX Low qPCR kit (Kapa Biosystems, Wilmington, MA, USA).…”

Section: Real-time Pcrmentioning

confidence: 99%

“…Quantitative PCR (qPCR) assays were carried out using the ViiA7 Real-Time PCR System and KAPA SYBER FAST ROX Low qPCR kit (Kapa Biosystems, Wilmington, MA, USA). 12 Relative gene expression levels were quantified by qPCR followed by normalization to the internal control gene 36B4. The following primers were used for this analysis: Kbtbd11 Fwd, 5 0 -TCAGCGTTTTCCGCTACCAT-3 0 and Kbtbd11 Rv, 5 0 -AACA CAACGAAAGGGCTGGA-3 0 ; Cebpa Fwd, 5 0 -GCCATGCCGG GAGAACTCTA-3 0 and Cebpa Rv, 5 0 -GGGCTCTGGAGGTG ACTGCT-3 0 ; Cebpb Fwd, 5 0 -GAGCCGCGACAAGGCCAAGA-3 0 and Cebpb Rv, 5 0 -GCTCGTTCTCCGCCGTCAGC-3 0 ; Pparg Fwd, 5 0 -TTCCACTATGGAGTTCATGCTTGT-3 0 and Pparg Rv, 5 0 -TCCGGCAGTTAAGATCACACCTA-3 0 ; p27 Fwd, 5 0 -AGGAGAGCCAGGATGTCAGC-3 0 and p27 Rv, 5 0 -CAGAGT TTGCCTGAGACCCAA-3 0 ; Cyclin D1 Fwd, 5 0 -GTTCATTTC CAACCCACCCTC-3 0 and Cyclin D1 Rv, 5 0 -AGAAAGTGCG TTGTGCGGTAG-3 0 ; Bax Fwd, 5 0 -GCTGACATGTTTGCT GATGG-3 0 and Bax Rv, 5 0 -GATCAGCTCGGGCACTTTAG-3 0 ; Bcl2 Fwd, 5 0 -CTGGGATGCCTTTGTGGAAC-3 0 and Bcl2 Rv, 5 0 -GAGACAGCCAGGAGAAATCAAAC-3 0 ; Tnfa Fwd, 5 0 -CAG CCGATGGGTTGTACCTT-3 0 and Tnfa Rv, 5 0 -GGGCTCA TACCAGGGTTTGA-3 0 ; Il6 Fwd, 5 0 -GAGGATACCACTCCC AACAGACC-3 0 and Il6 Rv, 5 0 -AAGTGCATCATCGTTGTTC ATACA-3 0 ; 36B4 Fwd, 5 0 -ATGCAGCAGATCCGCATGT-3 0 and 36B4 Rv, 5 0 -TTGCGCATCATGGTGTTCTT-3 0 .…”

Section: Real-time Pcrmentioning

confidence: 99%

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Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

Watanabe

Yoshida

Iwamoto

2019

J of Diabetes Invest

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Aims/Introduction The putative tumor suppressor gene, KBTBD11 , might play a role in tumorigenesis, and is associated with cellular apoptosis and proliferation in colorectal cancer cells. However, the function of Kbtbd11 during adipogenesis is unknown. The aim of the present study was to investigate the role of Kbtbd11 in the differentiation of 3T3‐L1 preadipocytes. Materials and Methods For the fasting–refeeding protocol, mice were subjected to fasting for 24 h, followed by a chow diet for 12 h. Adenovirus infection methods were used to examine the effect of Kbtbd11 , and 3T3‐L1 cells were analyzed with Oil Red O staining and real‐time polymerase chain reaction. Results The white adipose tissue expression of Kbtbd11 messenger ribonucleic acid (mRNA) was significantly higher in the re‐fed state than in the fasted state. Kbtbd11 mRNA levels were markedly increased in epididymal white adipose tissue of diet‐induced obesity mice compared with those in the mice fed a chow diet. In addition, Kbtbd11 mRNA expression was increased in a differentiation‐dependent manner in 3T3‐L1 cells. Knockdown of Kbtbd11 mRNA through the infection with adenoviral vectors remarkably inhibited triglyceride accumulation and adipocyte differentiation in 3T3‐L1 cells. In contrast, the overexpression of Kbtbd11 promoted the differentiation of 3T3‐L1 adipocytes. Conclusions The present findings show that Kbtbd11 expression might be involved in nutritional regulation and is increased in obese adipose tissue. In addition, Kbtbd11 appears to be required for the differentiation of adipocytes in 3T3 ‐ L1 cells. Collectively, these results show a novel link between the expression of Kbtbd11 and fat accumulation, and suggest that Kbtbd11 is a new therapeutic target for obesity.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Real-time Pcrmentioning

confidence: 99%

Section: Real-time Pcrmentioning

confidence: 99%

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Kbtbd11 gene expression in adipose tissue increases in response to feeding and affects adipocyte differentiation

Watanabe

Yoshida

Iwamoto

2019

J of Diabetes Invest

Self Cite

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show abstract

“…It has also been suggested that ILDR2, together with its homologue ILDR1, regulates alternative pre-mRNA splicing through binding to certain splicing factors (10). ZNF70, a member of the Kruppel C2H2-type zinc finger protein family, has been reported to bind to ILDR2 resulting in HES1 gene expression changes (11). However, these studies have been performed only in ILDR2-transfected cells in vitro.…”

Section: Introductionmentioning

confidence: 99%