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 | ||
| 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) |
| RT-qPCR | ||
| 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 | |
| SiRNA923 | Sense | 5′-CACCGAACGTGGTCAATGTATAATTCAAGAGATTATACATTGACCACGTTCTTTTTTG-3′ |
| Antisense | 5′-GATCCAAAAAAGAACGTGGTCAATGRATA | |
| ATCTCTTGAATTATACATTGACCACGTTC-3′ | ||
| Scrambled siRNA | Sense | 5′-CACCGTTCTCCGAACGTGTCACGTCAAGA |
| GATTACGTGACACGTTCGGAGAATTTTTG-3′ | ||
| Antisense | 5′-GATCCAAAAAATTCTCCGAACGTGTCA | |
| GTCTCTTGACGTGACACGTTCGGAGAAC-3′ | ||