Table 1.
Primers for the panel of genes used to measure relative gene expression
| Gene | Name | Forward/reverse primer sequences () | Reference |
|---|---|---|---|
| ACTB | actin β | F: CACAGATCATGTTTGAGACCTT R: CATCACAATACCAGTGGTACG | 12 |
| GAPDH | glyceraldehyde 3-phosphate dehydrogenase | F: GGCACGCCATCACTATC R: CCTGCATCTGCCCATTT | 12 |
| ANGPTL4 | angiopoietin-like 4 | F: TCCTCGATTCGCGAGTTCTG R: CAGGGCACTGGGAGCTG | 12 |
| CD72 | cluster of differentiation 72 | F: AGGAAGGTAGGGCAGCAATG R: CTGACCTGAGGTTCGCCAAA | 14 |
| CXCR5 | C-X-C motif chemokine receptor 5 | F: GCTCTGACTGTAGGGTGACG R: TGAAATGATGGGCAGTGGCT | 14 |
| CYR61 | cysteine-rich angiogenic inducer 61 | F: ATCGCTCGTTCAGACGCATA R: TGTCTGGGCTCCGCTAAAAG | 12 |
| IKZF1 | Ikaros family zinc finger 1 | F: GCGTGTGAAAGAGCGACTTC R: GAACACTCCGCACAACACCT | 14 |
| KLHL6 | kelch-like family member 6 | F: ATGGTTTCTGCGTCAACTCC R: CATCCTGGCTGGGATGCAATA | 12 |
| NR4A3 | nuclear receptor subfamily 4 group A member 3 | F: GGCATCCCCGGAGTTTCTCTG R: TTTGACGAGGCCGCTCATT | 12 |
| SERPING1 | serpin family G member 1 | F: GTCCTCGTGCCACACTTACC R: TTGACCAATGCTTGCCCACC | 14 |
| SYK | spleen-associated tyrosine kinase | F: AAGGGACAGCAATGGTTCCT R: AATTTAACAGACCTGCCAGAGG | 12 |
| TNFRSF14 | tumour necrosis factor receptor superfamily member 14 | F: TGAGCACCATCAGGGGTATC R: AGGTACGGATGCTTCCCAAG | 14 |
Table 2.
Primers for the panel of genes used for DNA methylation analysis
| Gene | Name | Methylated (M)/unmethylated (U) Forward (F)/Reverse (R) primer sequences | Reference |
|---|---|---|---|
| ANGPTL4 | angiopoietin-like 4 | MF: TAATTTTAACGGGAAGTATTTTCGT MR: CAACTTTAAAACTCTACCTCCAACG UF: TAATTTTAATGGGAAGTATTTTTGT UR: ACTTTAAAACTCTACCTCCAACACA | 12 |
| CD72 | cluster of differentiation 72 | MF: AACGGGTTATGTGTCGTTATTAGTC MR: AAACTAAACCCTACTACCTTCTCGC UF: TGGGTTATGTGTTGTTATTAGTTGT UR: ACTAAACCCTACTACCTTCTCACA | 12 |
| CXCR5 | C-X-C motif chemokine receptor 5 | MF: AGAGGTTGGGATTTACGGTAATAAC MR: ACAACTTTCTACCTTTACAAACGCT UF: AGGTTGGGATTTATGGTAATAATGT UR: ACAACTTTCTACCTTTACAAACACT | 12 |
| CYR61 | cysteine-rich angiogenic inducer 61 | MF: TTTGGTTTTAGTGTTTAAAGACGT MR: TTATATTTACCTTCAAAAAAACGTA UF: TTTTGGTTTTAGTGTTTAAAGATGT UR: TATTTATATTTACCTTCAAAAAAACATA | 12 |
| IKZF1 | Ikaros family zinc finger 1 | MF: GTAGTAGTAATTGTTGGAGGAGGC MR: AAAAATAACTTTACGAAACAACGAA UF: GTAGTAGTAATTGTTGGAGGAGGTG OR: AAAAATAACTTTACAAAACAACAAA | 12 |
| KLHL6 | kelch-like family member 6 | MF: TTTTTTGGATAATGAGTGTTTAACG MR: AAACACCAAAAAAAATCCCGTA UF: TTTTTGGATAATGAGTGTTTAATGA UR: CTAAAACACCAAAAAAAATCCCATA | 12 |
| NR4A3 | nuclear receptor subfamily 4 group A member 3 | MF: GGGAAAGGATAAAGTTTTTGTAGTC MR: AAACTCAAACGTAACCCTAAACGTA UF: GGGAAAGGATAAAGTTTTTGTAGTTG UR: AAACTCAAACATAACCCTAAACATA | 12 |
| SERPING1 | serpin family G member 1 | MF: GGTAACGAGAGTTTGGATTTGTAAC MR: CCTAAATAAACCCTAAAAACTACGC UF: TGGTAATGAGAGTTTGGATTTGTAAT UR: CTAAATAAACCCTAAAAACTACACC | 12 |
| SYK | spleen-associated tyrosine kinase | MF: TATTAGGCGTTTTCGGGAAC MR: AAATTAATACATTTACTCGCCGCT UF: GTTTATTAGGTGTTTTTGGGAATGA UR: CCAAATTAATACATTTACTCACCACT | 12 |
| TNFRSF14 | tumour necrosis factor receptor superfamily member 14 | MF: GTTTTAGTTATTTTTGTTTTTACGTTCGT MR: CCGCTATCACTATACAACTTCTCG UF: AGTTATTTTTGTTTTTATGTTTGT UR: CACTATCACTATACAACTTCTCACC | 12 |
Fig. 1.
Relative expression of selected genes in the liver of 42-day-old Ross 308 broilers after in-ovo sodium butyrate administration on day 12 of incubation. Values above 1 indicate upregulation, and values below 1 indicate downregulation compared to the control. FC – fold change; 0.1% SB – group administered 0.1% sodium butyrate; 0.3% SB – group administered 0.3% sodium butyrate; 0.5% SB – group administered 0.5% sodium butyrate; ANGPTL4 – angiopoietin-like 4; CD72 – cluster of differentiation 72; CXCR5 – C-X-C chemokine receptor type 5; CYR61 – cysteine-rich angiogenic inducer 61; IKZF1 – Ikaros transcription factor; KLHL6 – kelch-like family member 6; NR4A3 – nuclear receptor subfamily 4 group A member 3; SYK – spleen-associated tyrosine kinase; SERPING1 – serpin family G member 1. Data are presented as mean SEM (n = 8). Exact P-values shown above the brackets indicate significant differences between specific treatment groups (P-value < 0.05)
Fig. 2.
Extent of DNA methylation of selected genes in the liver of 42-day-old Ross 308 broilers after in-ovo sodium butyrate administration on day 12 of incubation. 0.1% SB – group administered 0.1% sodium butyrate; 0.3% SB – group administered 0.3% sodium butyrate; 0.5% SB – group administered 0.5% sodium butyrate; ANGPTL4 – angiopoietin-like 4; CD72 – cluster of differentiation; CXCR5 – C-X-C chemokine receptor type 5; CYR61 – cysteine-rich angiogenic inducer 61; IKZF1 – Ikaros transcription factor; KLHL6 – kelch-like family member; NR4A3 – nuclear receptor subfamily 4 group A member 3; SERPING1 – serpin family G member 1; SYK – spleen-associated tyrosine kinase; TNFRSF14 – tumour necrosis factor receptor superfamily member 14. Data are presented as mean and SEM (n = 8). Exact P-values shown above the brackets indicate significant differences between specific treatment groups (P-value < 0.05)
Fig. 3.
Global DNA methylation levels in the liver of 42-day-old Ross 308 broilers after in-ovo sodium butyrate administration on day 12 of incubation. 5-mC – 5-methylcytosine; 0.1% SB – group administered 0.1% sodium butyrate; 0.3% SB – group administered 0.3% sodium butyrate; 0.5% SB – group administered 0.5% sodium butyrate. Data are presented as mean and SEM (n = 8). No significant effect was noted on global DNA methylation (P-value = 0.1441)