Fig 1: PPARA regulated NCOR1 and protected damaged heart cells. (A) The JASPAR database was used to predict the presence of PPARA binding motifs in the promoter region of NCOR1. (B) ChIP assay was used to validate that the transcription factor PPARA was bound to the NCOR1 promoter region. After AC16s were transfected with oe-PPARA + si-NCOR1 and its negative control, the expressions of PPARA and NCOR1 were detected using (C) Western blot. Cell viability of each group was detected using (D) CCK-8 assay. The apoptosis rates of each group were detected using (E,F) flow cytometry. The contents of ANP, BNP, IL-1β and IL-18 were detected using (G) ELISA in cell supernatant. The expressions of Myh6 and Myh7 were detected by (H) Western blot. (I) ROS production, MDA production, and SOD activity were detected in each group of cells. (J) Mitochondrial ATP production was detected by the kit in cells. Compared with the Control group, *P<0.05; compared with the oe-NC + si-NC group, #P<0.05; compared with the oe-PPARA + si-NC group, &P<0.05. n=3. PPARA, peroxisome proliferator-activated receptor α; NCOR1, nuclear receptor co-repressor 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; DOX, doxorubicin; oe-NC, overexpression negative control; si-NC, small interfering RNA negative control; HCMEC, human cardiac microvascular endothelial cell; ChIP, chromatin immunoprecipitation; CCK-8, Cell Counting Kit-8; ELISA, enzyme-linked immunosorbent assay; ANP, atrial natriuretic peptide; BNP, brain natriuretic peptide; IL-1β, interleukin-1β; IL-18, interleukin-18; Myh6, myosin heavy chain 6; Myh7, myosin heavy chain 7; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase; ATP, adenosine triphosphate.
Fig 2: SIRT1 targeted the regulation of PPARA and affected heart cells. The expressions of SIRT1 and PPARA were detected by (A) Western blot in each group. Apoptosis rates were detected by (B,C) flow cytometry in each group. The contents of ANP, BNP, IL-1β and IL-18 were detected by (D,E) ELISA in the cell supernatant of each group. The expression of HF marker molecules Myh6 and Myh7 were detected using (F) Western blot. (G) ROS production, MDA production, and SOD activity were detected in each group of cells. (H) Mitochondrial ATP production was detected using a mitochondrial respiratory chain kit. Compared with the control group, *P<0.05; compared with the oe-NC + si-NC group, #P<0.05; compared with the oe-SIRT1 + si-NC group, &P<0.05. n=3. SIRT1, Sirtuin 1; PPARA, peroxisome proliferator-activated receptor α; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; DOX, doxorubicin; oe-NC, overexpression negative control; si-NC, small interfering RNA negative control; HCMEC, human cardiac microvascular endothelial cell; ELISA, enzyme-linked immunosorbent assay; ANP, atrial natriuretic peptide; BNP, brain natriuretic peptide; IL-1β, interleukin-1β; IL-18, interleukin-18; HF, heart failure; Myh6, myosin heavy chain 6; Myh7, myosin heavy chain 7; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase; ATP, adenosine triphosphate.
Fig 3: SIRT1 protected damaged heart cells. SIRT1 expression was detected by (A) RT-qPCR and (B) Western blot in AC16 and HCMEC cells. (C,D) Apoptosis rates were detected using flow cytometry. (E,F) The contents of ANP, BNP, IL-1β, and IL-18 were detected using ELISA in each group. (G) ROS production, MDA production and SOD activity were measured in each group. Compared with the control group, *P<0.05; compared with the oe-NC group, #P<0.05. n=3. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; HCMEC, human cardiac microvascular endothelial cell; DOX, doxorubicin; oe-NC, overexpression negative control; SIRT1, Sirtuin 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ELISA, enzyme-linked immunosorbent assay; ANP, atrial natriuretic peptide; BNP, brain natriuretic peptide; IL-1β, interleukin-1β; IL-18, interleukin-18; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase.
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