Total RNA was extracted from CA1 region of hippocampal (Fig. 1b), or cultured HT22 cells (Figs. 1d, 2a, 3a, 4a, 4b, 5a, 6a and 7A) by using TRIzol reagent (Life Technologies Corporation, Carlsbad, CA, USA). RNA concentration and quality of each sample were determined with Nanodrop Spectrophotometer (ND-100) by the 260/280 nm ratio. The primers for Snhg8, Hoxa13, and β-actin were synthesized from Takara Bio Inc. The expression levels of miR-384 and U6 (Applied Biosystems, Foster City, CA, USA) were examined with High Capacity cDNA Reverse Transcription Kits (Applied Biosystems, Foster City, CA, USA) and Taqman Universal Master Mix II (Life Technologies Corporation, Carlsbad, CA, USA).
Snhg8: forward 5′-GACACAAGGTGGCTATGGTGCTG-3′,
reverse 5′-CATGGTGGTCGTCGCGCTAAC-3′;
Hoxa13: forward5′-TACTTCGGCAGCGGCTACTACC-3′,
reverse 5′-CGGCGGTGTCCATGTACTTGTC-3′;
FAM3A: forward 5′-TTGGCCTTCCTCGAATTCAGCAG-3′,
reverse 5′-GCTCAGGTGCTCTTCAGGACAAG-3′;
β-actin: forward 5′-GTGACGTTGACATCCGTAAAGA-3′,
reverse 5′-GTAACAGTCCGCCTAGAAGCAC-3′.
The expression levels of miR-384 and U6 (Applied Biosystems, Foster City, CA, USA) were examined with High Capacity cDNA Reverse Transcription Kits (Applied Biosystems, Foster City, CA, USA) and Taqman UniversalMaster Mix II (Life Technologies Corporation, Carlsbad, CA, USA). The relative quantification 2−△△Ct method was applied to calculate the gene expression.
Liu J., An P., Xue Y., Che D., Liu X., Zheng J., Liu Y., Yang C., Li Z, & Yu B. (2019). Mechanism of Snhg8/miR-384/Hoxa13/FAM3A axis regulating neuronal apoptosis in ischemic mice model. Cell Death & Disease, 10(6), 441.
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