Background
Results
The human fibroblast cell lines HS 633T and HT-1080 are differently susceptible to CHIKV infection
Robust HS 633T- and milder HT-1080- innate immune responses against CHIKV
CHIKV interferes with the nuclear translocation of pSTAT1 in both HS 633T and HT-1080 cell lines
Discussion
Conclusion
Methods
Cells and virus
Infection protocol
Immunofluorescence
Quantitative real-time RT-PCR (CHIKV E1 gene)
Quantitative real time RT-PCR for innate immune genes
Reference | Gene | Primer | 5' modification | Sequence (5' to 3') | 3' modification |
---|---|---|---|---|---|
Primer and probes used for the one-step qRT-PCR from TAKARA
| |||||
NM_002201.4 |
ISG20
| ISG20-636 F | CTGTTGTGGCGTGAGGCCA | ||
ISG20-764R | TGCCCTCGCATCTTCCACC | ||||
ISG20-688P | HEX | TGCTGAGTGAGCGCCTCCTGC | BHQ-1 | ||
NM_002201.4 |
ISG54
| ISG54-76 F | GGTCTCTTCAGCATTTATTGGTG | ||
ISG54-219R | TGCCGTAGGCTGCTCTCCA | ||||
ISG54-124P | FAM | TGCAGCTGCCTGAACCGAGC | BHQ-1 | ||
NM_001548.3 |
ISG56
| ISG56-131 F | GCCTAATTTACAGCAACCATGAG | ||
ISG56-347R | GGCCTTTCAGGTGTTTCACATA | ||||
ISG56-215P | HEX | TGGGAGTTATCCATTGATGACGATG | BHQ-1 | ||
NM_014314.3 |
RIG-I
| RIG-I-237 F | AGCTACATGGCCCCCTGGT | ||
RIG-I-397R | GCATGGTCTAGGGCATCCAA | ||||
RIG-I-351P | HEX | CAGGAGGAAGGCTGGTTCCGT | BHQ-1 | ||
NM_002046 |
GAPDH
| GAPDH-F | AGCCTCAAGATCATCAGCAATG | ||
GAPDH-R | CTGTGGTCATGAGTCCTTCCA | ||||
GAPDH-P | HEX | CCAACTGCTTAGCACCCCTGGC | BHQ1 | ||
Primers used for the RT-qPCR with BRYT Green
| |||||
NM_002176.2 |
IFN-β
| F | GTTCGTGTTGTCAACATGACCA | ||
R | TCAATTGCCACAGGAGCTTCT | ||||
NM_016562.3 |
TLR7
| F | CCACAACCAACTGACCACTG | ||
R | CCACCAGACAAACCACACAG | ||||
NM_002046.3 |
GAPDH
| F | GCACCGTCAAGGCTGAGAAC | ||
R | GCCTTCTCCATGGTGGTGAA |