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Jessica Miciak

on 2 April 2014

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Transcript of NCATS Trial

Transfections of A204
Using Fugene 6
Getting a free sample of Lipofectamine 3000
Wnt/TGF-B Cross Talk
Jess's Data - April 2, 2014
A204 Brightfield 48hrs Transfection
pSmad3 Staining
Try Lipofectamine 3000 for C2C12 transfections
Do in parallel with 293T Cells
Next Steps
Open Field testing
Treadmill Exhaustion (+ Open Field)
Cardiac Testing
Muscle Harvesting/Histology
Open Field #2 (7pm-9pm)
Open Field Following Exhaustion on Treadmill
Open Field 1st Trial - Tested 9am-4pm
Sectioning FSHD muscle biopsies
Staining with specific DSHB antibodies
Waiting on Myosin IIx antibody to confirm differential staining of type II fibers
FSHD Fiber Typing
Four Major Fiber Types - "slow" I, "fast" IIA, IIB, IIX
Speed of Contraction
Sarcoplasmic reticulum development
Resistance to fatique
Mitochondrial content
Thickness of z-line
In Human MyHC-2B not detectable
Fibers typed 2B based on ATPase staining are 2x fiber based on MyHC composition
Combined Staining
Fiber Typing - Separate
Treadmill Exhaustion
p = 0.245
p = 0.166
p = 0.062
-Con- -Mstn- -TGFB-
2 hour treatment C2C12
Full transcript