ShRNAs against METTL3 (sh-METTL3#1, sh-METTL3#2, sh-METTL3#3), NAT10 (sh-NAT10#1, sh-NAT10#2, sh-NAT10#3), and their negative controls (sh-NC) were provided by Shanghai GenePharma Company (China). NAT10 was overexpressed by subcloning its entire sequence into the pcDNA3.1 vector to construct the overexpression vector (pcDNA3.1-NAT10), with the pcDNA3.1 empty vector used as a control. TE-5 and Eca-109 cells seeded in eight-well plates were transfected with 80 nM of synthesized shRNAs using Lipofectamine 3000 (Invitrogen, USA) and incubated for 48 h.
Sh nat10
Manufactured by GenePharma
Sourced in China
Sh-NAT10 is a laboratory equipment product developed by GenePharma. It is designed for the purpose of small hairpin RNA (shRNA) expression. The core function of Sh-NAT10 is to facilitate the stable expression of shRNA in mammalian cell lines.
Automatically generated - may contain errors
3 protocols using sh nat10
1
Knockdown and Overexpression of METTL3 and NAT10
2
NAT10 Knockdown and Overexpression
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Plasmids for NAT10 knockdown (sh-NAT10) and overexpression (OE-NAT10) were synthesized directly (GenePharma, Shanghai, China). sh-NAT10 and OE-NAT10 vectors were delivered via lentiviral infection into SW480 and DLD-1 cells. Furthermore, cells that were infected using lentiviruses with scrambled plasmids were utilized as negative controls. After a span of 48 h, the cells were collected in order to conduct additional studies.
3
Overexpression and Knockdown of NAT10 and KLF6 in HGFs
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The pcDNA3.1-NAT10 and pcDNA3.1-KLF6 expression plasmids were constructed by Shanghai Biological Engineering Company. HGFs were seeded in six-well plates (2 × 105 cells / well) prior to transfection. The transfection was performed using Effectene Transfection Reagent (QIAGEN Companies) according to the manufacturer’s instructions when cell confluence reached to over 60% confluence. The experimental and control groups were termed pc-NAT10, pcKLF6 and pcDNA3.1, respectively.
As for NAT10 inhibition, the short hairpin RNA (shRNA) lentivirus of NAT10 (shNAT10) and negative control (shNC) lentivirus were constructed by GenePharma Co., LTD. HGFs were transfected with the specified vector using the Lipofectamine 3000 (Invitrogen) regeant for 48 h.
As for NAT10 inhibition, the short hairpin RNA (shRNA) lentivirus of NAT10 (shNAT10) and negative control (shNC) lentivirus were constructed by GenePharma Co., LTD. HGFs were transfected with the specified vector using the Lipofectamine 3000 (Invitrogen) regeant for 48 h.
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