Essential role of ATP6AP2 enrichment in caveolae/lipid raft microdomains for the induction of neuronal differentiation of stem cells

Table 2 Distribution of TUJ1⁺/O4⁺/GFAP⁺ purified cells before and after hADSC differentiation, and their relative mRNA ATP6AP2 expression

	Immunophenotyping^a (% of total cells)		ATP6AP2 mRNA^b (fold expression relative to MSCs)
	MSCs Day < 0	NLCs Day 37	NLCs Day 37
Healthy
TUJ1⁽⁺⁾	2.87 ± 0.14	76.04 ± 0.22 ^***	18.2 ± 0.49 ^***
O4⁽⁺⁾	5.10 ± 0.38	21.12 ± 0.53 ^**	4.27 ± 0.80 ^*
GFAP⁽⁺⁾	20.92 ± 0.46	0.91 ± 0.65 ^***	1.31 ± 0.42
ND
TUJ1⁽⁺⁾	1.94 ± 0.90	31.44 ± 0.95 ^{**, §§}	6.11 ± 0.83 ^{*, §§}
O4⁽⁺⁾	1.08 ± 0.67	10.08 ± 0.48 ^{*, §}	5.09 ± 0.78 ^*
GFAP⁽⁺⁾	32.16 ± 0.71 ^§	8.23 ± 0.51 ^{*, §}	1.07 ± 0.95

hADSC human adipose-derived mesenchymal stem cell, MSC mesenchymal stem cell, ND neurodegenerative disease, NLC neural-like cell, NSP neurosphere-like structure
^a2 × 10⁷ cells derived each from healthy or ND subjects were applied to purification at day < 0 and day 37 as indicated in Methods. Three positive fractions were purified separately: TUJ1⁽⁺⁾, O4⁽⁺⁾ and GFAP⁽⁺⁾ cells. Cells were labeled with anti-TUJ1-FITC or anti-O4-APC or anti-GFAP-PE and analyzed using a MACSQuant flow analyzer as indicated in Methods. Results presented as mean ± SEM of two independent experiments each performed in duplicate
^bExpression of ATP6AP2 mRNA was determined by RT-qPCR on the purified fraction. Results expressed as fold variation relative to MSCs after normalization to GAPDH. Cells were collected at day 0 before the induction of the differentiation (MSCs) or at day 37 of the differentiation (NLCs)
^*P < 0.05, ^**P < 0.01 and ^***P < 0.005: NLCs vs MSCs
^§P < 0.05, ^§§P < 0.01: ND vs healthy

ISSN: 1757-6512