Si-RNA and Lentivirus Transfection

ADGRG1−targeting small interfering RNAs (si-RNAs) and negative control siRNAs (si−NC) were designed and synthesized by GenePharmaCorp. The ADGRG−siRNA sequences were as follows: siRNA-1428, 5’−GCAACCACUUGACCUACUUTT−3’; siRNA-1900, 5’−GGUGGAUGUGGACAACUAUTT−3’; siRNA-2178, 5’−CCUUUGCUUCUGGCACCU UTT−3’. The si−NC sequence was as follows: 5’−UUCUCCGAACGUGUCACGUTT−3’ (sense) and 5’−ACGUGACACGUUCGGAGA ATT−3’ (antisense). The si−RNAs and si−NC were transfected into Siha by INTERFERin (Polyplus Transfection) according to the manufacturer’s protocols. After 48 h, knockdown efficiency was evaluated by PCR and Western blotting. Lentiviruses carrying short hairpin RNA (shRNA: 5′‐GCAACCACTTGACCTACTT-3′) targeting ADGRG1 and negative control (shNC: 5′‐TTCTCCGAACGTGTCACGT-3′) were also synthesized by GenePharmaCorp.