Si00193025

Example 5

Hif-1a siRNA Transfection In Vitro and In Vivo.

B6, SW, and MRL ear fibroblast-like cells at 70% of confluence were transfected with 100 nM of 4 different HIF-1a siRNAs (SI00193025, SI00193032, SI00193011, SI00193018) purchased from Qiagen and scramble siRNA (sc-37007, Santa Cruz Biotechnologies), using Lipofectamine 2000 according to the manufacturer's protocol. Transfected cells were examined for the knockdown efficiency after 48 h of transfection. siRNA Mm_Hif1a_3 (SI00193025) was selected for the in vitro experiments due to its high efficiency. In vivo, siRNA Mm_Hif1a_3 was used for HIF-1a inhibition. SiHif at 75 mg/kg body weight was mixed with Jetpei (Polyplus, Genycell) following manufacturer's instructions and was then injected into animals subcutaneously every 48 h.

Example 5

Hif-1a siRNA Transfection In Vitro and In Vivo.

B6, SW, and MRL ear fibroblast-like cells at 70% of confluence were transfected with 100 nM of 4 different HIF-1a siRNAs (SI00193025, SI00193032, Sl00193011, SI00193018) purchased from Qiagen and scramble siRNA (sc-37007, Santa Cruz Biotechnologies), using Lipofectamine 2000 according to the manufacturer's protocol. Transfected cells were examined for the knockdown efficiency after 48 h of transfection. siRNA Mm_Hif1a_3 (SI00193025) was selected for the in vitro experiments due to its high efficiency. In vivo, siRNA Mm_Hif1a_3 was used for HIF-1a inhibition. SiHif at 75 mg/kg body weight was mixed with Jetpei (Polyplus, Genycell) following manufacturer's instructions and was then injected into animals subcutaneously every 48 h.