fabp4 sirna  (Santa Cruz Biotechnology)

Journal: Experimental and Therapeutic Medicine

Article Title: FABP4 alleviates endoplasmic reticulum stress-mediated ischemia-reperfusion injury in PC12 cells via regulation of PPARγ

doi: 10.3892/etm.2021.9612

Figure Lengend Snippet: Downregulation of FABP4 in PC12 cells under OGD/R conditions. (A) PC12 cells were maintained under OGD for 2, 4, 6, 8 or 10 h followed by reoxygenation for 24 h. Cell viability was estimated using a Cell Counting Kit-8 assay. (B) mRNA expression of FABP4 in PC12 cells exposed to OGD/R were determined by RT-qPCR. * P<0.05, ** P<0.01, *** P<0.001 vs. control. The (C) protein and (D) mRNA expression of FABP4 in PC12 cells were determined by RT-qPCR and western blotting following transfection with FABP4 siRNA or control siRNA. ** P<0.01, *** P<0.001 vs. si-Con. (E) The protein expression of FABP4 in PC12 cells exposed to OGD 8 h/R (OGD for 8 h and reoxygenation for 24 h) was determined by western blotting. *** P<0.001 vs. control; ### P<0.001 vs. OGD 8 h/R+si-Con. FABP4, fatty acid binding protein 4; siFABP4, FABP4 small interfering RNA; siCon, control small interfering RNA; OGD/R, oxygen glucose deprivation/reoxygenation; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Cells were transfected with 10 μM FABP4 small interfering RNA (siRNA; Santa Cruz Biotechnology, Inc.) or negative control siRNA using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol.

Techniques: Cell Counting, Expressing, Quantitative RT-PCR, Control, Western Blot, Transfection, Binding Assay, Small Interfering RNA, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Experimental and Therapeutic Medicine

Article Title: FABP4 alleviates endoplasmic reticulum stress-mediated ischemia-reperfusion injury in PC12 cells via regulation of PPARγ

doi: 10.3892/etm.2021.9612

Figure Lengend Snippet: FABP4-knockdown inhibits oxidative stress and apoptosis. (A) Cell proliferation was measured by Cell Counting Kit-8 assay. (B) ROS production and (C) SOD activity were assessed using a ROS assay kit (magnification, x100) or SOD assay kit, respectively. (D) Cell apoptosis was detected by flow cytometry. (E) Cleaved caspase-3, caspase-3 and β-actin were estimated by western blotting. ** P<0.01, *** 0.001 vs. control; # P<0.05, ## P<0.01, ### P<0.001 vs. OGD 8 h/R+si-Con. FABP4, fatty acid binding protein 4; ROS, reactive oxygen species; SOD, superoxide dismutase; siCon, control small interfering RNA; OGD/R, oxygen glucose deprivation/reoxygenation.

Article Snippet: Cells were transfected with 10 μM FABP4 small interfering RNA (siRNA; Santa Cruz Biotechnology, Inc.) or negative control siRNA using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol.

Techniques: Knockdown, Cell Counting, Activity Assay, ROS Assay, Flow Cytometry, Western Blot, Control, Binding Assay, Small Interfering RNA

Journal: Experimental and Therapeutic Medicine

Article Title: FABP4 alleviates endoplasmic reticulum stress-mediated ischemia-reperfusion injury in PC12 cells via regulation of PPARγ

doi: 10.3892/etm.2021.9612

Figure Lengend Snippet: Interference of FABP4 suppresses inflammation and ERS and activates PPARγ. (A) GRP78, CHOP and β-actin expression was estimated by western blotting. (B) mRNA expression of TNF-α, IL-1β and IL-6 were determined by reverse transcription-quantitative polymerase chain reaction. (C) Protein expression of PPARγ was detected by western blotting. ** P<0.01, *** 0.001 vs. control; # P<0.05, ## P<0.01, ### P<0.001 vs. OGD 8 h/R+si-Con. FABP4, fatty acid binding protein 4; ERS, endoplasmic reticulum stress; PPARγ, peroxisome proliferator-activated receptor γ; GRP78, glucose regulating protein 78; CHOP, C/EBP homologus protein; TNF-α, tumor necrosis factor α; IL, interleukin; OGD/R, oxygen glucose deprivation/reoxygenation.

Article Snippet: Cells were transfected with 10 μM FABP4 small interfering RNA (siRNA; Santa Cruz Biotechnology, Inc.) or negative control siRNA using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol.

Techniques: Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Binding Assay

Journal: Experimental and Therapeutic Medicine

Article Title: FABP4 alleviates endoplasmic reticulum stress-mediated ischemia-reperfusion injury in PC12 cells via regulation of PPARγ

doi: 10.3892/etm.2021.9612

Figure Lengend Snippet: FABP4 modulates ERS and apoptosis via regulation of PPARγ. (A) PC12 cells exposed to OGD 8 h/R. Cells were pretreated with 1 µΜ GW9662 or 1 µg/ml TM for 2 h before OGD 8 h/R. GRP78, CHOP and β-actin expression was estimated by western blotting. (B) Cleaved caspase-3, caspase-3 and β-actin expression was estimated by western blotting. (C) Cell apoptosis was detected by flow cytometry. *** P<0.001 vs. control; ### P<0.001 vs. OGD 8 h/R; Δ P<0.05, ΔΔ P<0.01 vs. OGD 8 h/R+si-FABP4-1. FABP4, fatty acid binding protein 4; ERS, endoplasmic reticulum stress; PPARγ, peroxis(^#me proliferator-activated receptor γ; TM, tunicamycin; OGD/R, oxygen glucose deprivation/reoxygenation; GRP78, glucose regulating protein 78; CHOP, C/EBP homologous protein.

Article Snippet: Cells were transfected with 10 μM FABP4 small interfering RNA (siRNA; Santa Cruz Biotechnology, Inc.) or negative control siRNA using Lipofectamine 2000 (Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol.

Techniques: Expressing, Western Blot, Flow Cytometry, Control, Binding Assay