To silence Mdh2, mPDCs were transduced, in the presence of 8μg/ml polybrene (Sigma-Aldrich, Diegem, Belgium) with a lentivirus carrying the gene-targeting shRNA (Sigma-Aldrich) the day after isolation. A lentivirus containing an empty vector (Sigma-Aldrich) was used as negative control. To silence Tet2, cells isolated from Tet2fl/fl mice and wild-type (Tet2wt/wt) littermates were transduced with a lentivirus carrying a Puro.Cre empty vector (Addgene, Watertown, MA, USA). After 72h, puromycin (2μg/ml; InvivoGen, San Diego, CA, USA) was added to the culture medium for 48h. Sequences of the shRNAs are listed in Table S2 .
Lentivirus carrying
Manufactured by Merck Group
Sourced in Belgium
Lentivirus carrying is a tool used in research and laboratory settings. It is a modified virus that can be used to deliver genetic material into target cells. The core function of this product is to facilitate the introduction and expression of specific genes or genetic sequences within the recipient cells, enabling researchers to study gene function, protein expression, and other biological processes.
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2 protocols using lentivirus carrying
1
Silencing Mdh2 and Tet2 in mPDCs
2
Rat Ventricular Cardiomyocyte Isolation and Treatments
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Rat ventricular cardiomyocytes were isolated by enzymatic digestion from 1- to 3-day-old neonatal Sprague-Dawley rats and cultured as described previously [22] (link). The following treatments were pursued for the cell cultures, i.e., stimulation of cardiomyocytes with 1 µmol/L Ang II (human angiotensin II from Sigma, St Louis, MO); Lentivirus carrying miR-34a mimics (miR-34a), miR-34a inhibitor (miR-34a inhibitors), negative control of miR-34a, ATG9A siRNA, or negative control-siRNA, all having been purchased from the corporation of GenePharma, China (sequences are shown in Table 1 ). For lentiviral transductions, 4×105 of cardiomyocytes were transduced with 20 µl lentivirus (109 TU/ml) and polybrene (at a final concentration of 5 µg/ml). pRC/CMV2-ATG9A and vector control were transfected with Lipofectamine LTX and PLUS Reagents (Invitrogen) according to the manufacturer's instructions.
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