Undifferentiated N2aC, both WT and KO cell lines, were plated onto glass bottom video dishes and allowed to adhere for 24 hrs. After 24hrs, cells were incubated in EMEM media containing 0.5% fetal bovine serum and 20 μM retinoic acid (Millipore Sigma, St Louis, MO) for 24 hrs. After incubation in retinoic acid containing media, N2aC display a change in cell body morphology and extend MAP2 positive neurites, confirmed in live cells using an ZOE Fluorescent Cell Imager (Bio-Rad, Hercules, CA). After confirming neurite extension, cells were transfected with mechanistic target of rapamycin activity reporter (TORCAR) as described above.
Retinoic acid
Manufactured by Bio-Rad
Retinoic acid is a form of vitamin A used in various laboratory techniques. It is a key component in cell culture media, playing a role in cellular differentiation and development processes. Retinoic acid is a widely used reagent in biological research applications.
Automatically generated - may contain errors
2 protocols using retinoic acid
1
Neuronal Differentiation of N2aC Cells
2
Grafting Retinoic Acid and Shh Beads
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Sieved formate‐derivatized AG1‐X2 beads (150 or 200 μm in diameter, Sigma) were soaked in all‐trans‐retinoic acid (Sigma, 1 μg/μl‐1 or 5 μg/μl‐1 dissolved in DMSO, also Sigma) for 1 hr and then washed twice in DMEM before being grafted to the anterior margin of chick wing buds using a sharp tungsten needle. Affi‐Gel beads (Bio‐Rad) were soaked in recombinant Shh protein (10 μg/μl‐1, a kind gift of Prof Joy Richman) for 2 hr and implanted the same way as retinoic acid beads.
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