Table 1 Antibodies and sequences used for Western blotting and RT-qPCR. And sequences of siRNA923 and scrambled siRNA
Protein | Mfrs. /Made in | Cat No. |
|---|---|---|
Primary antibodies | ||
AFP | Santa Cruz BiotechInc./USA | sc-8399 |
PTEN | Cell Signaling Technology/USA | 9188S |
LC3 | Sigma/USA | L7543-100UL |
PI3K | Cell Signaling Technology/USA | 4249P |
AKT | Cell Signaling Technology/USA | 2920P |
mTOR | Cell Signaling Technology/USA | 2983P |
p-PI3K | Cell Signaling Technology/USA | 4228P |
p-AKT | Cell Signaling Technology/USA | 4060P |
p-mTOR | Cell Signaling Technology/USA | 5536P |
P62 | Cell Signaling Technology/USA | 5114S |
Flag | Sigma/USA | F7425 |
GAPDH | Sigma/USA | G9545 |
Secondary antibodies | Â | Â |
HRP-conjugated anti-mouse IgG | Zhongshan Boil Tech Co/China | IH-0031 |
HRP-conjugated anti- rabbitIgG | Zhongshan Boil Tech Co/China | IH-0011 |
Alexa Fluor 568 | Thermo Fisher Scientific/USA | A-11004 |
Alexa Fluor 488 | Thermo Fisher Scientific/USA | A-11034 |
Gene | Primer sequences | Product size (bp) |
| Â | RT-qPCR | Â |
P62 | Sense | 5′-GGTGGCACTTGTTATGCTATCCT-3'75 |
|  | Antisense 5′-GCCCATGGGAACTCACAATC-3′ |  |
AFP: | Sense | 5′-CCAACAGGAGGCCATGCTT-3'61 |
|  | Antisense | 5′-GAATGCAGGAGGGACATATGTTT-3′ |
GAPDH | Sense | 5′-TGAAGGTCGGAGTCAACGGA-3'232 |
|  | Antisense | 5′-CCTGGAAGATGGTGATGGGAT-3′ |
AFP | SiRNA | Â |
SiRNA923 | Sense | 5′-CACCGAACGTGGTCAATGTATAATTCAAGAGATTATACATTGACCACGTTCTTTTTTG-3′ |
|  | Antisense | 5′-GATCCAAAAAAGAACGTGGTCAATGRATA |
|  |  | ATCTCTTGAATTATACATTGACCACGTTC-3′ |
Scrambled siRNA | Sense | 5′-CACCGTTCTCCGAACGTGTCACGTCAAGA |
|  |  | GATTACGTGACACGTTCGGAGAATTTTTG-3′ |
|  | Antisense | 5′-GATCCAAAAAATTCTCCGAACGTGTCA |
|  |  | GTCTCTTGACGTGACACGTTCGGAGAAC-3′ |
