Mmu rnu6

Total microRNAs were extracted and purified from frozen mesenteric vessels by using miRNeasy Mini Kit (Qiagen, Germany). Reverse transcription of the extracted miRNAs was performed by the miScript Reverse Transcription Kit (Qiagen, Germany). cDNA was diluted 1:3 in RNase-free water and then qPCRs were performed in triplicate using the miScript SYBR-Green PCR kit (Qiagen, Germany) on the MiniOpticon CFX 48 real-time PCR Detection System (Bio-Rad, Hercules, CA, United States). MiScript Primer Assays specific for mmu-miR-214-3p (MIMAT0000661), mmu-SNORD6 and mmu-RNU6 were obtained from Qiagen. miR-214-3p expression was calculated using Ct method and normalized to the expression of two housekeeping, SNORD6 and RNU6.

Total microRNAs were purified and extracted from cryo-preserved colon (20–50 mg) and epididymal fat (100mg) by using miRNeasy Mini Kit (Qiagen, Germany). Reverse transcription of the extracted miRNAs was performed by the miScript Reverse Transcription Kit (Qiagen, Germany). cDNA was diluted 1:10 in RNase-free water and then qPCRs were performed in triplicate using the miScript SYBR-Green PCR kit (Qiagen, Germany) on the MiniOpticon CFX 48 real-time PCR Detection System (Bio-Rad, Hercules, USA). MiScript Primer Assays specific for mmu-let-7f-5p (MIMAT0000525) and the housekeeping mmu-RNU6 were obtained from Qiagen. let-7f expression was calculated using Ct method and normalized to the expression of RNU6.