Foxc2

Total RNA was extracted using RNeasy Mini Kit (Qiagen Inc., Valencia, CA, USA) and total RNA (1 μg) was synthesized with the ReverTra Ace qRT Kit (Toyobo, Osaka, Japan). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were performed on StepOne Plus Real-Time PCR Systems (Applied Biosystems, Foster City, CA, USA) using TaqMan Fast Universal PCR Master Mix (Applied Biosystems) and analyze the relative standard curve quantification method. PCR condition was according to the manufacturer’s instructions and GAPDH mRNA level was amplified for internal control. TaqMan Gene Expression Assays of Prox1, FOXC2, VEGF-A, VEGF-C, VEGF-D, and GAPDH were purchased from Applied Biosystems. All PCRs were done at triplicate.

Real-time PCR analysis was performed as previously described [24 (link),27 (link)]. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was utilized in experiments as a control gene [24 (link)]. We purchased the primers and probes for mouse Osterix, Osteocalcin, Osteopontin, Foxc2, Flk1, VE-cadherin, and CD31 from Applied Biosystems (Waltham, MA, USA) as part of Taqman^® Gene Expression Assays.