Total RNA from the hippocampus of post-COVID HM mice and controls was extracted using TRIzol reagent (Thermo Scientific, USA). cDNAs was synthesized and subjected to qPCR using Power SYBR Green Master Mix kits following the manufacturer’s instructions on the QuantStudio™ 7 Flex real-time PCR system platform (Applied Biosystems, USA). Primer sequences used are listed in Table 3 . Gene expression changes were determined by the 2–ΔCt method using ribosomal protein L32 for normalization.
Primer sequences for RT-qPCR.
| Gene Name | Forward primer (5’-3’) | Reverse primer (5’-3’) |
|---|---|---|
| IL-1β | TTGACGGACCCCAAAAGAT | GAAGCTGGATGCTCTCATCTG |
| IL6 | GATGGATGCTACCAAACTGGA | CCAGGTAGCTATGGTACTCCAGAA |
| TNF1 | TGCTGGGAAGCCTAAAAGG | CGAATTTTGAGAAGATGATCCTG |
| TNF2 (link) | GCTGAGCTCAAACCCTGGTA | CGGACTCCGCAAAGTCTAAG |
| IFNβ | GCCCTGTAGGTGAGGTTGATCT | AGCTCCAAGAAAGGACGAACAT |
| PSD-95–Dgl4 | TCTGTGCGAGAGGTAGCAGA | AAGCACTCCGTGAACTCCTG |
| BDNF | ATGAAAGAAGTAAACGTCCAC | CCAGCAGAAAGAGTAGAGGAG |
| RpL32 | GCTGCCATCTGTTTTACGG | TGACTGGTGCCTGATGAACT |
| MHV-3 | CAGATCCTTGATGATGGCGTAGT | AGAGTGTCCTATCCCGACTTTCTC |
1 Primer used for RT-PCR in lung and gut samples; 2 (link)Primer used for RT-PCR in brain samples; IL, interleukin; TNF, Tumor necrosis factor; IFN, interferon; PSD-95–Dgl4, Post-synaptic density protein; BDNF, Brain-derived neurotrophic factor; RpL, Ribosomal protein; MHV-3, Murine Hepatitis Virus-3 nucleocapsid protein (N) gene.
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