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Unless otherwise stated, glucose in the medium was substituted with 0.9 mg/ml galactose to force ATP production by the respiratory chain7.
When indicated, the 2% glucose of the defined medium was substituted by glycerol (in fact, 2% glycerol and 0.2% glucose).
The next day, the medium was substituted with DMEM/F12 containing 0.1% FBS with or without siRNA to render the cells quiescent.
On day 2 (d2), the serum containing medium was substituted with serum replacement medium (KO) and this resulted in initial proliferation and a gradual switching to differentiation.
The medium was substituted with Knockout-DMEM (Kon on d2 that contained all the additives and Knockout serum replacement (Invitrogen) in place of FBS and was supplemented with (KR) or without 100 nM all-trans retinoic acid (RA).
At these time points, culture medium was substituted by RHB-A: Neurobasal: B27 (1∶1∶0.02) medium supplemented either with 0.01% DMSO (control) or with 3 nM LY411575 (in 0.01% DMSO).
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When another medium is substituted, as in boards painted to look like stone, the result looks fake.
In order to initiate the maturation phase, the cytokinins and auxin in the liquid GD medium were substituted with 20 μM abscisic acid (ABA, Sigma-Aldrich) and 3.75% polyethylene glycol 4000 (PEG, Sigma-Aldrich).
Finally, higher total LDL-P and lower total HDL-P independently predicted CHD, adjusted for each other, age, race-ethnicity, BMI, WC, physical activity, diabetes, smoking, hypertension, lipid-lowering medication and hormone therapy (Table 6), with similar results if medium HDL-P was substituted for total HDL-P.
For EMM minus nitrogen supplemented with uracil medium (EMM-N+U), NH4Cl was substituted with 200 mg/liter of uracil.
Percoll gradient purification was used for epithelial isolation and was far more successful than using culture medium were L-Valine was substituted for D-Valine in the presence of which only cancer cells can continue grow.
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