BMDM or RAW 264.7 cells were transiently transfected with siRNA targeting STAT1 (75 nM), LXRα (100 nM), LXRβ (100 nM), or control siRNA (Bioneer, Seoul, Korea) using 5 μl of siRNA transfection reagent (Genlantis, San Diego, CA) according to the manufacturer’s protocol. The sequences used for STAT1 knockdown were sense 5′-CACAGUUUUAUCCUGAUGA-3′ and antisense 5′-UCAUCAGGAUAAAACUGUG-3′. The sequences used for LXRα knockdown were sense 5′-CGUAGCAUUAAGGGAGAGU-3′ and antisense 5′-ACUCUCCCUUAAUGCUACG-3′. The sequences used for LXRβ knockdown were sense 5′-ACGCUUACACCUCAGCCUA-3′ and antisense 5′-UAGGCUGAGGUGUGUAAGCGU-3′. The sequences for the control siRNA were sense 5′-CCUACGCCACCAAUUUCGU-3′ and antisense 5′-ACGAAAUUGGUGGCGUAGG-3′. The cells were then incubated in serum-free culture medium for 48 or 66 h.
Sirna transfection reagent
Manufactured by Genlantis
The SiRNA transfection reagent is a laboratory tool used for the delivery of small interfering RNA (siRNA) into cells. It facilitates the efficient introduction of siRNA molecules into the target cells, enabling the study of gene expression and function through RNA interference (RNAi) techniques.
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Lab products found in correlation
5 protocols using sirna transfection reagent
1
siRNA Knockdown of STAT1, LXRα, and LXRβ
2
Targeted Knockdown of COX-2, Axl, and Mer
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LA-4 cells were transiently transfected with 1 μg/mL of siRNA specifically targeting either COX-2, Axl, Mer or with control siRNA (Bioneer, Seoul, Korea) using 5 μL of siRNA transfection reagent (Genlantis, San Diego, CA, USA) according to the manufacturer’s protocol. The sequences used for COX-2 knockdown were sense 5′-CUAUGAUAGGAG CAUGUAA-3′ and antisense, 5′-UUACAU GCUCCUAUCAUA G-3′. The sequences used for Axl knockdown were sense 5′-GAGAUGGACA GAUCCUAGA-3′ and antisense, 5′-UCUAGGAUCUGUCCAUCUC-3′. The sequences used for Mer knockdown were sense 5′-CACAGUUUUAUCCUGAUGA-3′ and antisense 5′-UCAUCAGGAUAA AACUGUG-3′. The sequences for control siRNA were sense 5′-CCUACGCCACCAAUUUCG U-3′ and antisense 5′-ACGAAAUUGGUGGCGUAG G-3′. Cells were incubated in serum-free medium for 6 h for COX-2 siRNA, or 48 h for Axl and Mer siRNA prior to experimentation. None of the siRNAs used had any significant effect on cell viability.
3
RhoA, Axl, and Mer Knockdown in LA-4 Cells
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LA-4 cells were transiently transfected with 1 µg/mL of siRNA that specifically targeted either RhoA, Axl, or Mer or with control siRNA (Bioneer, Seoul, Korea), using 5 µL of siRNA transfection reagent (Genlantis, San Diego, CA, USA) according to the manufacturer’s protocol. The siRNA sequences used for targeting genes were as follows (gene: sense, antisense). RhoA (#1): 5′-GAAGUCAAGCAUUUCUGUCTT A-3′, 5′-GACAGAAAUGCUUGACUUCTT-3′; RhoA (#2): 5′-GUCUCAUGUUAGUUACCUUTT-3′, 5′-AAGGUAACUAACAUGAGACTT-3′; Axl (#1): 5′-GAGAUGGACAGAUCCUAGA-3′, 5′-UCUAGGAUCUGUCCAUCUC-3′; Axl (#2): 5′-CACACACUCAAGAAUCCAATT-3′, 5′-UUGGAUUCUUGAGUGUGUGTT-3′; Mer (#1): 5′-CACAGUUUUAUCCUGAUGA-3′, 5′-UCAUCAGGAUAAAACUGUG-3′; Mer (#2): 5′-UGACAGAAACCUUCUGGUUTT-3′, 5′-AACCAGAAGGUUUCUGUCATT-3′. Cells were incubated in serum-free medium for 24 h prior to siRNA experiments. None of the siRNAs used had any significant effects on cell viability.
4
Knockdown of COX-2, COX-1, and RhoA in Macrophages
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RAW 264.7 cells and resident peritoneal macrophages were transiently transfected with 1 μg/mL of either siRNA specifically targeting COX-2 or COX-1 or control siRNA (Bioneer, Seoul, Korea) using 5 μL of siRNA transfection reagent (Genlantis, San Diego, CA) according to the manufacturer's protocol. The sequences used for COX-2 knockdown were 5′-CUA UGA UAG GAG CAU GUA A-3′ (sense) and 5′-UUA CAU GCU CCU AUC AUA G-3′ (antisense). The sequences used for COX-1 knockdown were 5′-GAG GUA GGA ACU UUG ACU A-3′ (sense) and 5′-UAG UCA AAG UUC CUA CCU C-3′ (antisense). The sequences for control siRNA were 5′-CCU ACG CCA CCA AUU UCG U-3′ (sense) and 5′-ACG AAA UUG GUG GCG UAG G-3′ (antisense). Before further experiments, cells were incubated in serum-free medium for 6 h for COX-2 siRNA or 48 h for COX-1 siRNA. For RhoA siRNA, RAW 264.7 cells were transiently transfected with 10 nM RhoA-targeting siRNA (sense: 5′-GAA GUC AAG CAU UUC UGU CTT-3′; antisense: 5′-GAC AGA AAU GCU UGA CUU CTT-3′) premixed with 6 μg/mL of Lipofectin (Invitrogen). Cells were then incubated in serum-free medium for 24 h before further experimentation. None of the siRNAs used had any significant effect on cell viability.
5
Knockdown of COX-2, COX-1, and RhoA in RAW 264.7 Cells
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RAW 264.7 cells were transiently transfected with 1 μg/ml of siRNA specifically targeting either COX-2, COX-1, or RhoA or with control siRNA (Bioneer, Seoul, Korea) using 5 μl of siRNA transfection reagent (Genlantis, San Diego, CA, USA) according to the manufacturer’s protocol. The sequences used for COX-2 knockdown were as follows: sense, 5′-CUA UGA UAG GAG CAU GUA A-3′; and antisense, 5′-UUA CAU GCU CCU AUC AUA G-3′. The sequences used for COX-1 knockdown were as follows: sense, 5′-GAG GUA GGA ACU UUG ACU A-3′; and antisense, 5′-UAG UCA AAG UUC CUA CCU C-3′. The sequences used for RhoA knockdown were as follows: sense, 5′-GAA GUC AAG CAU UUC UGU CTT A-3′; and antisense, 5′- GAC AGA AAU GCU UGA CUU CTT-3′. The sequences for control siRNA were as follows: sense, 5′-CCU ACG CCA CCA AUU UCG U-3′; and antisense, 5′-ACG AAA UUG GUG GCG UAG G-3′. Cells were incubated in serum-free medium for 6 h for COX-2 siRNA, 48 h for COX-1 siRNA, or 24 h for RhoA siRNA prior to experimentation. None of the siRNAs used had any significant effect on cell viability.
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