Table 1
List of primer sequences for quantitative real-time polymerase chain reaction
| Gene | Primer sequence (5′→3′) | Amplicon size | |
|---|---|---|---|
| Bax | Forward: | TGCAGAGGATGATTGCTGAC | 173 bp |
| Reverse: | GATCAGCTCGGGCACTTTAG | ||
| Casp3 | Forward: | CTGCCGGAGTCTGACTGGAA | 97 bp |
| Reverse: | ATCAGTCCCACTGTCTGTCTCAATG | ||
| Trp73 (p73) | Forward: | AGCCTTTGGTTGACTCCTATCG | 119 bp |
| Reverse: | ACCACCGTGTACCTTGTTCA | ||
| Ccna2 | Forward: | ACCTGCCTTCACTCATTGCT | 139 bp |
| Reverse: | TGGTGAAGGTCCACAAGACA | ||
| Ccnb1 | Forward: | CAGTTGTGTGCCCAAGAAGA | 216 bp |
| Reverse: | CTACGGAGGAAGTGCAGAGG | ||
| Ccne1 | Forward: | CACCACTGAGTGCTCCAGAA | 230 bp |
| Reverse: | CTGTTGGCTGACAGTGGAGA | ||
| Cdkn1a | Forward: | GGGATGGCAGTTAGGACTCA | 244 bp |
| Reverse: | GTGGGGCAAGTGCCTAGATA | ||
| Cdk4 | Forward: | CAATGTTGTACGGCTGATGG | 178 bp |
| Reverse: | CAGGCCGCTTAGAAACTGAC | ||
| Rps6 | Forward: | GTCCGCCAGTATGTTGTCAG | 103 bp |
| Reverse: | GTTGCAGGACACGAGGAGTA | ||
| Kitl | Forward: | TCATGGTGCACCGTATCCTA | 170 bp |
| Reverse: | CCTTGGCATGTTCTTCCACT | ||
| Bdnf | Forward: | GGTATCCAAAGGCCAACTGA | 183 bp |
| Reverse: | CTTATGAATCGCCAGCCAAT | ||
| Ntrk2 (TrkB) | Forward: | CGCCCTGTGAGCTGAACTCTG | 171 bp |
| Reverse: | CTGCTTCTCAGCTGCCTGACC | ||
| Ntrk1 (TrkA) | Forward: | GAGGTCTCTGTCCAAGTCAGCG | 131 bp |
| Reverse: | TTGAACAACCAGCGCAGAGA | ||
| Ngf | Forward: | CAGACCCGCAACATCACTGTA | 131 bp |
| Reverse: | CCATGGGCCTGGAAGTCTAG | ||
| Tet1 | Forward: | CGAAAGAACAGCCACCAGAT | 219 bp |
| Reverse: | TTGCTCTTCTTCCCCATGAC | ||
| Tet2 | Forward: | GTTGCAAGAAGAAAGCGGAG | 229 bp |
| Reverse: | CTCTGCCCTTGCTGAAGGT | ||
| Tet3 | Forward: | TCCGGATTGAGAAGGTCATC | 176 bp |
| Reverse: | CCAGGCCAGGATCAAGATAA | ||
| Dnmt1 | Forward: | CCTAGTTCCGTGGCTACGAGGAGAA | 136 bp |
| Reverse: | TCTCTCTCCTCTGCAGCCGACTCA | ||
| Dnmt3a | Forward: | CACAGGGCCCGTTACTTCTG | 76 bp |
| Reverse: | TCCAGCTTATCATTCACAGTGGAT | ||
| Dnmt3b | Forward: | TTCAGTGACCAGTCCTCAGACACGAA | 144 bp |
| Reverse: | TCAGAAGGCTGGAGACCTCCCTCTT | ||
| Mecp2_e1 | Forward: | AGGAGAGACTGGAGGAAAAGT | 70 bp |
| Reverse: | CTTAAACTTCAGTGGCTTGTCT | ||
| Mecp2_e2 | Forward: | CTCCATAAAAATACAGACTCACCAGT | 183 bp |
| Reverse: | CTTAAACTTCAGTGGCTTGTCT | ||
| β-actin | Forward: | ATGGTGGGAATGGGTCAGAAG | 157 bp |
| Reverse: | CACGCAGCTCATTGTAGAAGG |
Figure 1
Cell viability of mHypoE-N46 cells following PFOA treatment for (A) 24 h and (B) 48 h (MTT assay). Data are shown as mean ± standard error of triplicate measurements. ** p<0.01 and *** p<0.001 compared to control
Figure 2
Expression of (A) apoptosis-related genes, (B) cell cycle-related genes, and (C) cell proliferation-related genes after 24 h of exposure to PFOA (qPCR). Data are shown as mean ± standard error of three independent measurements. * p<0.05, ** p<0.01, and *** p<0.001 compared to control
Figure 3
Expression of (A) ligand (Ngf) and receptor (Ntrk1) genes of the NGF-NTRK1 neurotrophin system and (B) ligand (Bdnf) and receptor (Ntrk2) genes of the BDNF-NTRK2 neurotrophin system after 24 h of exposure to PFOA (PCR). Data are shown as mean ± standard error of three independent measurements. * p<0.05, ** p<0.01, and *** p<0.001 compared to control
Figure 4
Epigenetic toxicity of PFOA: (A) relative global methylation levels; (B) relative expression of ten-eleven translocation methylcytosine dioxygenase-encoding Tet1, Tet2, and Tet3; (C) relative expression of DNA methyltransferase-encoding Dnmt1, Dnmt3a, and Dnmt3b, and (D) relative expression of methyl-CpG binding protein 2 genes. Data are shown as mean ± standard error of three independent measurements. * p<0.05, ** p<0.01, and *** p<0.001 compared to control