To silence LUZP1 and EPLIN, RPE-1 cells (105 cells seeded in 6-well plates) were transfected with 40 nM (final concentration) of a pool of two siRNAs targeting either gene obtained from Dharmacon (ON-TARGET plus) using the Lipofectamine RNAiMAX transfection reagent (Invitrogen) according to the manufacturer’s instructions. The Luciferase GL2 Duplex nontargeting siRNA from Dharmacon was used as a negative control. Gene silencing was performed for 72 h. The siRNA sequences are listed in Table S2 . When indicated, the cells were serum starved (DMEM/F12 supplemented with GlutaMAX and sodium bicarbonate [1.2 g/liter] but without serum) for 72 h to induce the formation of primary cilia.
Luciferase gl2 duplex
Manufactured by Horizon Discovery
The Luciferase GL2 Duplex is a laboratory instrument used for the detection and measurement of luciferase enzyme activity. Luciferase is a bioluminescent enzyme commonly used as a reporter in various biological applications. The Luciferase GL2 Duplex provides a reliable and efficient method for quantifying luciferase activity in samples.
Automatically generated - may contain errors
2 protocols using luciferase gl2 duplex
1
Silencing LUZP1 and EPLIN in RPE-1 Cells
2
Transfection of siRNAs in Cells
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The following siRNAs were used in this study: Luciferase (siLuc, 20 or 100 nM, Luciferase GL2 duplex; Dharmacon/D-001 100–01-20), Bub1 (siBub1, 100 nM, 5′-GAAUGUAAGCGUUCACGAADTDT-3′; Dharmacon), WAPL (siWAPL, 40 nM, 5′-GAGAGAUGUUUACGAGUUUDTDT-3′; Dharmacon), and Haspin (siGSG2, 40 nM, Ambion AM51331, ID1093). Cells were plated and transfected immediately using HiPerfect (Qiagen) or DharmaFECT 2 (Dharmacon), according to the manufacturers’ instructions.
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