HaCaT cells were purchased from Pricell (Wuhan, China) and cultured with RPMI 1640 medium (BasalMedia, Shanghai, China) containing 10% FBS and 1% penicillin/streptomycin. After reaching 60% confluence, the cells were stimulated with 5.5 mmol/L glucose or 35 mmol/L glucose for 48 h. Then cells were transfected with 50 nmol/L miRNA mimic NC or miR-221-3p mimic (GenePharma, Shanghai, China), and 50 nmol/L small interfering RNA (siRNA) sequence 1 and 2 of DYRK1A or siRNA NC (GenePharma) for another 48 h using lipofectamine 3000 (L3000015, Thermo Fisher Scientific, USA). The medium was changed every 2 days. HaCaT cell supernatant was collected to measure cytokine content and the cells were harvested for subsequent analysis. The siRNA sequences used in this study are shown in Supplementary Table 5 .
Mirna mimic nc
Manufactured by GenePharma
Sourced in China
The MiRNA mimic NC is a laboratory equipment product designed for microRNA (miRNA) studies. Its core function is to provide a non-coding miRNA control sequence for use in miRNA-related experiments and analyses. The product enables researchers to establish baseline measurements and controls within their miRNA research projects.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using mirna mimic nc
1
Glucose-induced miR-221-3p Modulation of DYRK1A
2
Modulating Macrophage Responses via Exosomal miRNA and siRNA
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RAW264.7 cells were seeded in 6-well plates to 60–80% confluence and divided into four groups. Two groups were transfected with 10 µl of 20 nm miR-155-5p mimics and mimic-normal control (miRNA mimic-NC), and miR-155-5p inhibitors and miRNA inhibitor-normal control (miRNA inhibitor-NC) (Gene Pharma, Suzhou, China), respectively. The other two groups were transfected with 10 µl of 20 nm si-socs1 (siRNA targeting socs1) and si-NC (siRNA-NC) (RiboBio, Guangzhou, China), respectively. The transfection was performed using Lipofectamine 3000 (Thermo Fisher Scientific) according to the protocol provided by the manufacturer. The exosomes extracted from the cell culture supernatant of the DC2.4 (DC-Exo) and the DC2.4 infected with T. gondii (Tg-DC-Exo) were added to the 60–80% confluent RAW264.7 cells at 120 µg/well, respectively.
3
Silencing lncRP5 using shRNAs and miR-545-5p modulation
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Short hairpin RNAs (shRNAs) against lncRP5 which were named as sh-LNCRP5#1 and sh-lncRP5#2, were used to silence lncRP5 in cells and the negative control was denoted as sh-NC. miR-545-5p mimics, inhibitors, and the corresponding negative controls (miRNA NC) were synthesized by Biosyn Biotech Co., Ltd (Suzhou, China), and the sequences are as follows: sh-lncRP5#1 sense 5′-ACCTCGCGAGAGAGATTCCGATTTATTCAAGAGATAAATCGGAATCTCTCTCGCTT-3′and antisense 5′-CAAAAAGCGAGAGAGATTCCGATTTATCTCTTGAATAAATCGGAATCTCTCTCGCG-3′; sh-lncRP5#2 sense 5′-ACCTCGACCAAACAGGACCTTTACCTTCAAGAGAGGTAAAGGTCCTGTTTGGTCTT-3′ and antisense 5′-CAAAAAGACCAAACAGGACCTTTACCTCTCTTGAAGGTAAAGGTCCTGTTTGGTCG-3′; sh-NC 5′-GTTCTCCGAACGTGTCACGTCAAGAGATTACGTGACACGTTCGGAGAATT-3′; miR-545-5p mimics sense 5′-UCAGCAAACAUUUAUUGUGUGC-3′ and antisense 5′-GCACACAAUAAAUGUUUGCUGA-3′; miR-545-5p inhibitor 5′-GCACACAAUAAAUGUUUGCUGA-3′; miRNA inhibitor NC 5′-CAGUACUUUUGUGUAGUACAA-3′; miRNA mimic NC sense 5′-UUCUCCGAACGUGUCACGUTT-3′ and antisense 5′-ACGUGACACGUUCGGAGAATT-3′. The whole sequences of lncRP5 and PTP4A1 were cloned into pcDNA3.1 vector (Genepharma, Shanghai, China) and transfected into the cells. The empty vectors were set as control. Cells were transfected with the small RNAs or the plasmid using Lipofectamine 3000 Reagent (Invitrogen) according to the manufacturer’s instructions.
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