qRT-PCR
Splenocytes or corneal tissues were lysed using RNA isolation reagent
(RNA Bee, Tel-Test, Friendswood, TX) and homogenized with an ultrasound
sonicator (Ultrasonic Processor, Cole Parmer Instruments, Vernon Hills,
IL). Total RNA was extracted using RNeasy Mini kit (Qiagen, Hilden,
Germany), and first-strand cDNA was synthesized by reverse transcription
(High Capacity RNA-to-cDNATM Kit, Applied Biosystems,
Carlsbad, CA). Real-time amplification was performed using
TaqMan® Universal PCR Master Mix (Applied Biosystems)
on an automated instrument (ABI 7500 Real Time PCR System, Applied
Biosystems) for Fut1 or Thbs1 . The data were normalized toGapdh and expressed as fold changes relative to controls. All PCR
probe sets were purchased from Applied Biosystems
(TaqMan® Gene Expression Assay kits, Applied
Biosystems).