Table 1 Reagents and primers details
Antibodies used for immunocytochemistry Antibody | Dilution | Cat # and RRID |
|---|---|---|
Pluripotency markers | ||
Mouse anti-OCT4 | 1:200 | CST Cat.75463 |
Rabbit anti-NANOG | 1:200 | CST Cat.4903 T |
Mouse anti-TRA1-60 | 1:500 | Abcam Cat.ab/6288 |
Mouse anti-SSEA4 | 1:500 | CST Cat.MC813 |
NSC differentiation markers | ||
Mouse anti-Nestin | 1:200 | ABcam.Cat.ab18102 |
Rabbit anti-sox1 | 1:200 | CST Cat.4194 s |
Secondary antibodies | ||
Mouse anti-IgG | 1:1000 | CST Cat.4409 |
Goat-anti-Rabbit | 1:500 | Servicebio. Cat.GB25303 |
Primers | Target | Forward/Reverse primer (5′-3′) | |
|---|---|---|---|
Targeted mutation | ALDH5A1-Exon5 | ACGTGACTTTAGCACTAATAAGA | AGAGCTTTTAACACTCTGCTGGA |
RT-qPCR Primers | ALDH5A1-RNA-1 (exons 8–10) | GGCACCAGTTATCAAGTTCG | ACTCCACCAAAAGGGCACTC |
ALDH5A1-RNA-2 (exons 3–4) | AGTCATCACCCCGTGGAATTT | GAGAAGGGCGTGTCTTCGG | |
ALDH5A1-RNA-3 (exons 5–6) | AGGGGAGGCAATTTGTACTGA | GTGGTGCAACAGGATCTTTCC | |
GAPDH | GCACCGTCAAGGCTGAGAAC | TGGTGAAGACGCCAGTGG | |
β-ACTIN | CATGTACGTTGCTATCCAGGC | CTCCTTAATGTCACGCACGAT | |
cDNA sequencing | F1/R1 | TTCCTGTCGCCGTCGTTGC | ATCCTACAAGCCCCCGTAACAC |
F2-1 | GGTCCTCAAGCAGCCCATA | ||
F2-2 | TTTGTAAAAGCATTCGCCG | ||
F3 | GATGCCGTTTCTAAAGGTGC | ||
R2 | AAAATAACCTGCTAACCCAACATC | ||
R3 | TTACAAGGACTGGATGAGTTCT | ||