Tb green fast qpcr kit

A549 cells (1 × 10⁶) were seeded in 6-well plates and treated with HBSS for 24 h. The cells were lysed using TRIzol buffer, and transcriptome sequencing and analysis was performed by Oebiotech (Shanghai, China). Real-time quantitative reverse transcription polymerase chain reaction was performed to confirm the transcriptional level of ATG proteins. Briefly, the cells were treated with rapamycin, lysed with TRIzol buffer, and reverse-transcribed using a PrimeScript™ RT reagent kit with gDNA Eraser (Takara Bio). Real-time polymerase chain reaction (PCR) was then performed using a TB Green Fast qPCR kit (Takara Bio). The primers used in this study were as follows: ATG3, forward: 5′-GAC​CCC​GGT​CCT​CAA​GGA​A-3′, reverse: 5′-TGTAGCCC ATTGCCATGTTGG-3'; ATG5, forward: 5′-AAA​GAT​GTG​CTT​CGA​GAT​GTG​T-3′, reverse: 5′-CAC​TTT​GTC​AGT​TAC​CAA​CGT​CA-3'; ATG7, forward: 5′-ATG​ATC​CCT​GTA​ACT​TAG​CCC​A-3′, reverse: 5′-CAC​GGA​AGC​AAA​CAA​CTT CAAC-3'; LC3B, forward: 5′-GAT​GTC​CGA​CTT​ATT​CGA​GAG​C-3'; reverse: 5′-TTG​AGC​TGT​AAG​CGC​CTT​CTA-3'; Beclin-1, forward: 5′-CCA​TGC​AGG​TGA​GCT​TCG​T-3′, reverse: 5′-GAA​TCT​GCG​AGA​GAC​ACC​ATC-3'; SQSTM1, forward: 5′-GCA​CCC​CAA​TGT​GAT​CTG​C-3′, reverse: 5′-CGCTACACAAGTC GTAGTCTGG-3'.