MDA-MB-231 cells were cultured in DMEM (HyClone; Cytiva) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO2. Subsequently, MDA-MB-231 cells (5×104 cells/well; 24-well plates) were transfected with 100 nM inhibitor control (cat. no. MIH000000; Applied Biological Materials, Inc.), 100 nM miR-1297 inhibitor (cat. no. MIH01244; Applied Biological Materials, Inc.), 1 µg control-plasmid (cat. no. sc-437275; Santa Cruz Biotechnology, Inc.), 1 µg FA2H-plasmid (cat. no. sc-413143-ACT; Santa Cruz Biotechnology, Inc.), 0.2 µM control-small interfering (si)RNA (cat. no. sc-36869; Santa Cruz Biotechnology, Inc.), 0.2 µM FA2H-siRNA (cat. no. sc-93418; Santa Cruz Biotechnology, Inc.), miR-1297 inhibitor + control-siRNA or miR-1297 inhibitor + FA2H-siRNA using Polyplus transfection reagent (Polyplus-transfection SA) according to the manufacturer's protocol. At 48 h post-transfection, cells were used for subsequent experiments.
Fa2h sirna
Manufactured by Santa Cruz Biotechnology
Sourced in United States
FA2H siRNA is a small interfering RNA (siRNA) molecule designed to target the FA2H gene, which encodes the enzyme fatty acid 2-hydroxylase. This siRNA can be used to suppress the expression of the FA2H gene in cells, allowing researchers to study the biological functions of this enzyme.
Automatically generated - may contain errors
Lab products found in correlation
Control sirna, by Santa Cruz Biotechnology (2 mentions)
Control plasmid, by Santa Cruz Biotechnology (1 mentions)
Control small interfering si rna, by Santa Cruz Biotechnology (1 mentions)
Hyclone, by Cytiva (1 mentions)
Dp22 digital camera, by Olympus (1 mentions)
Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (1 mentions)
Ckx41 inverted microscope, by Olympus (1 mentions)
3 protocols using fa2h sirna
1
MDA-MB-231 Cell Transfection Optimization
2
FA2H Regulation in AGS Cells
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AGS cells were transfected with 1 µg control plasmid (Cat no. sc-437275; Santa Cruz Biotechnology), 1 µg FA2H lasmid (Cat no. sc-413143-ACT; Santa Cruz Biotechnology), 0.2 µM control small interfering RNA (siRNA; Cat no. sc-36869; Santa Cruz Biotechnology), 0.2 µM FA2H siRNA (Cat no. sc-93418; Santa Cruz Biotechnology), 100 nM inhibitor control (5′-CAGUACUUUUGUGUAGUACAA-3′; GenePharma, Shanghai, China), 100 nM miR-300 inhibitor (5′-AGAGAGAGUCUGCCUUGUAUA-3′; GenePharma), 100 nM miR-300 inhibitor + 0.2 µM control siRNA or 100 nM miR-300 inhibitor + 0.2 µM FA2H siRNA for 48 h with Polyplus transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturers’ instructions before subsequent experimentation.
3
Cell Migration Assay for MDA-MB-231 Cells
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The cancer cell wound-healing assay was performed as described previously, with minor modifications (Takeda et al., 2012; Hirao-Suzuki et al., 2020a) . Briefly, MDA-MB-231 cells were seeded into 6-well plates at a density of approximately 3 × 10 5 cells/well and incubated for 4 hr; the cells were then treated with PFOA. Twenty-four hours after PFOA treatment, FA2H siRNA (sc-93418; Santa Cruz Biotechnology, Dallas, TX, USA) and Control siRNA (sc-37007; Santa Cruz Biotechnology, Inc.) were transfected using Lipofectamine™ RNAiMAX reagent (Thermo Fisher Scientific). Forty-eight hours after PFOA treatment (i.e., 24 hr after siRNA transfection), the monolayers were scratched with plastic tip. To visualize cell migration into the scratched region, the cells were imaged using an Olympus CKX41 inverted microscope (Tokyo, Japan) and an Olympus DP22 digital camera connected to a DP2-SAL camera controller after incubation for 0 hr, 12 hr, and 24 hr (i.e., treatment with PFOA for 48 hr, 60 hr and 72 hr, respectively) (Supplemental Fig. 1B).
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