Table 2.
Reagents details.
| Antibodies used for immunocytochemistry
| |||
|---|---|---|---|
| Antibody | Dilution | Company Cat# and RRID | |
| Pluripotency markers | Goat anti-NANOG | 1:400 | R and D Systems Cat# AF1997 RRID:AB_355097 |
| Pluripotency markers | Rabbit anti-OCT4 | 1:250 | Santa Cruz Biotechnology Cat# sc-9081 RRID:AB_2167703 |
| Pluripotency markers | Mouse anti-SSEA4 | 1:200 | R and D Systems FA1435P-025 |
| Pluripotency markers | Mouse anti-TRA-1-85 | 1:400 | R and D Systems Cat# FAB3195A RRID:AB_663789 |
| Differentiation markers | Anti-CD105 | 1:400 | Thermo Fisher Scientific Cat# 12-1057-42 RRID:AB_1311123 |
| Differentiation markers | Anti-CD73 | 1:400 | BD Biosciences Cat# 550257 RRID:AB_393561 |
| Differentiation markers | Rabbit anti-HNF4a |
1:1000 | Cell Signaling Technology Cat# 3113S RRID:AB_2295208 |
| Differentiation markers | Goat anti-SOX17 | 1:500 | R and D Systems Cat# AF1924 RRID:AB_355060 |
| Differentiation markers | Rabbit anti-PAX6 | 1:200 | BioLegend Cat# 901301 RRID:AB_2565003 |
| Differentiation markers | Mouse anti-NESTIN | 1:200 | BioLegend Cat# 655102 RRID:AB_2562023 |
| RB1 (Western Blot) | Mouse anti-RB1 | 1:250 | BD Biosciences Cat# 554136 RRID:AB_395259 |
| Secondary antibodies | Goat anti-rabbit IgG(Alexa Fluor 488 conjugate) | 1:1000 | Jackson ImmunoResearch Labs Cat# 111-545-144 RRID:AB_2338052 |
| Secondary antibodies | Donkey Anti-Goat IgG (Alexa Fluor488 conjugate) | 1:1000 | Jackson ImmunoResearch Labs Cat# 705-545-003 RRID:AB_2340428 |
| Secondary antibodies | Donkey Anti-Mouse IgG (Alexa Fluor488 conjugate) | 1:1000 | Jackson ImmunoResearch Labs Cat# 715-545-150 RRID:AB_2340846 |
| Secondary antibodies | Donkey Anti-Rabbit IgG (Cy3 conjugate) | 1:1000 | Jackson ImmunoResearch Labs Cat# 711-165-152 RRID:AB_2307443 |
| Secondary antibodies | Donkey Anti-Rabbit IgG (Cy5 conjugate) | 1:1000 | Jackson ImmunoResearch Labs Cat# 711-175-152 RRID:AB_2340607 |
| Primers | |||
|
| |||
| Target | Forward/reverse primer (5′–3′) | ||
|
| |||
| Pluripotency markers (qPCR) | OCT4 | AACCTGGAGTTTGTGCCAGGGTTT/TGAACTTCACCTTCCCTCCAACCA | |
| Pluripotency markers (qPCR) | SOX2 | AGAAGAGGAGAGAGAAAGAAAGGGAGAGA/GAGAGAGGCAAACTGGAATCAGGATCAAA | |
| Pluripotency markers (qPCR) | NANOG | TTTGTGGGCCTGAAGAAAACT/AGGGCTGTCCTGAATAAGCAG | |
| Pluripotency markers (qPCR) | DPPA4 | GACCTCCACAGAGAAGTCGAG/TGCCTTTTTCTTAGGGCAGAG | |
| Pluripotency markers (qPCR) | REX1 | GCCTTATGTGATGGCTATGTGT/ACCCCTTATGACGCATTCTATGT | |
| House-keeping genes (qPCR) | GAPDH | CCACTCCTCCACCTTTGAC/ACCCTGTTGCTGTAGCCA | |
| Targeted mutation analysis/sequencing | RB1 exon16 | TTCTTTTTATAGAAGTAAGTATTTTATAATC/CTCAAAGGTCTTCGGAGGGA | |