Table 1.
Primers used for RT-PCR.
| Gene | Primer Sequence (5’-3’ Product Size | Reference | |
|---|---|---|---|
| β-actin | F: AGATCAAGATCATCGCGCCT | 170 bp | Xia et al. (2017) |
| R: ATGCAACTAACAGTCCGCCT | |||
| IL-6 | F:CCTCGGCAAAATCTCTGCAA | 189 bp | Xia et al. (2017) |
| R: TGAAACTCCACAAGACCGGT | |||
| IL-8 | F: CCTCATTCCTGTGCTGGTCA | 273 bp | Xia et al. (2017) |
| R: TGCAAGTTGAGGCAAGAAGAC | |||
| TNF-α | F: GCCCTTCCACCAACGTTTTC | 158 bp | Xia et al. (2017) |
| R: TCCCAGGTAGATGGGTTCGT | |||
| pBD-1 | F: CTCCTCCTTGTATTCCTCCT | 141 bp | Wan et al. (2013) |
| R: GGTGCCGATCTGTTTCAT | |||
| pBD-2 | F: GACTGTCTGCCTCCTCTC | 148 bp | Wan et al. (2013) |
| R: GGTCCCTTCAATCTGTTG | |||
| IL-22 | F: CCCAGATCTGGGTACCATGGTCCCG | 477bp | primer premier 5.0 |
| R: GCCTTTAATACGACATTGGGACAGTT | |||
Supplementary Fig. 1 verification of rIL-22. (A) Identification of pET32a-IL-22 and double-enzyme digestion. lane 1 shows the recombinant plasmid of pET32a-IL-22; lane 2 shows the products after pET32a-IL-22 was digested by Kpn I and Hind III, Lane 3 shows the DNA marker; (B) Verification of colony PCR. Lane1 shows the DNA marker; lane 2 shows negative control; lane 3–10 show positive colony. (C) Western blot analysis of expressed recombinant rIL-22. Lane M2: Western blot marker; Lane 3: Supernatant of cell lysate with induction for 16 h at 15 °C; Lane 4: Pellet of cell lysate with induction for 16 h at 15 °C; Lane 5: Supernatant of cell lysate with induction for 4 h at 37 °C; Lane 6: Pellet of cell lysate with induction for 4 h at 37 °C.