Transfection was performed using cells in the logarithmic growth phase. The cells are seeded into six-well plates at a density of 5 × 105 cells/well. After the cells grow to 60% of the area of each well, transfection is performed. The specific procedure was carried out with Lipofectamine 3,000 according to the manufacturer’s instructions (Invitrogen, Thermo Scientific, MA, United States). The shRNA targeting TTN-AS1 (shTTN-AS1) and the shRNA targeting scramble (shSCR) were obtained from Shanghai GenePharma Co., Ltd (Shanghai, China).
Sh ttn as1
Manufactured by GenePharma
Sourced in China
Sh-TTN-AS1 is a laboratory equipment product. It is a short hairpin RNA (shRNA) targeting the titin (TTN) gene. The core function of Sh-TTN-AS1 is to modulate the expression of the TTN gene in experimental settings.
Automatically generated - may contain errors
3 protocols using sh ttn as1
1
Transfection of TTN-AS1 knockdown
2
Knockdown of TTN-AS1 and SFN using shRNAs
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For knockdown of TTN-AS1 or SFN, specific short hairpin RNAs (sh-RNAs) against TTN-AS1 (sh-TTN-AS1) or SFN (sh-SFN) were produced by Genepharma (Shanghai, China) and nonspecific sh-RNAs were used as the negative controls. The oligonucleotides of miR-513a-5p mimic and inhibitor, as well as the corresponding negative control NC mimic and inhibitor, were also acquired from Genepharma. The indicated plasmids and oligonucleotides were transfected into cells using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.
3
Knockdown of lncRNA TTN-AS1 in A549 cells
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Lentivirus vectors containing short hairpin RNA (shRNA) against TTN-AS1 (sh-TTN-AS1) or negative control (sh-con) were constructed by GenePharma (Shanghai, China). When cell confluence reached 70%, 1×106 TU/mL lentivirus supplemented with polybrene were infected into A549 cells. Next, puromycin was used to select stable cell clones.
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