NFκB1在奶牛乳腺上皮细胞中的表达与定位

doi:10.16431/j.cnki.1671-7236.2016.08.025

Abstract

Abstract:

The experiment was aimed to study the expression and localization of NFκB1 in bovine mammary epithelial cell (BMEC),and to clarify the regulation of transcription for milk synthesis in BMEC by NFκB1.Primary cells were cultured using the tissue pieces culture method,and the BMEC was purified and passaged according to the different sensitivity of fibroblasts and BMEC to trypsin.Western blotting and immunofluorescence (IF) method were used to detect cell keratin 18 and β-casein expression to identify the cell purity and lactogenesis function.Furthermore,0.6 mmol/L methionine (Met) was added to BMEC culture to establish amino acids-stimulating BMEC model in vitro.Western blotting and IF were used to detect the expression and localization of NFκB1 and p-NFκB1 after adding Met.These results showed that BMEC was successfully purified.NFκB1 had two molecular forms in cytoplasm,the molecular weights were about 141 and 105 ku,and in nucleus only 105 ku form exists.The p-NFκB1 (50 ku) mainly existed in the nucleus.After Met stimulation,the protein level of NFκB1 141 ku form was slightly upregulated,the protein level of NFκB1 105 ku did not changed in the cytoplasm while obviously increased in the nucleus,the protein level and nuclear localization of p-NFκB1 were both obviously increased.These results suggested that NFκB1 was involved in the transcriptional regulation of lactogenesis in BMEC,and amino acids promoted lactogenesis by triggering NFκB1 phosphorylation in the nucleus.

Key words: NFκB1; p-NFκB1; BMEC; methionine; lactogenesis

CLC Number:

S814.8

ZANG Yan-li, GU Xin-yu, HUANG Xin, YU Cui-ping, LUO Chao-chao, GAO Xue-jun. Expression and Localization of NFκB1 in Bovine Mammary Epithelial Cell[J]. , 2016, 43(8): 2112-2119.

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Expression and Localization of NFκB1 in Bovine Mammary Epithelial Cell

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