Journal: Frontiers in molecular biosciences

Article Title: Circular RNA TLK1 Promotes Sepsis-Associated Acute Kidney Injury by Regulating Inflammation and Oxidative Stress Through miR-106a-5p/HMGB1 Axis.

doi: 10.3389/fmolb.2021.660269

Figure Lengend Snippet: FIGURE 2 | Depletion of circTLK1 decreases oxidation stress, inflammation, and apoptosis in the sepsis-related AKI rat model. (A–G) The CLP-induced sepsis- related AKI rat model was established in Wistar rats (N 4), followed by the treatment of circTLK1 shRNA. The cecum was exposed without perforation in the sham groups. (A) MDA levels in the rats. (B) ROS production in the rats. (C) GSH levels in the rats. (D) CAT activities in the rats. (E) SOD activities in the rats. (F) IL-1β and IL-6 release in the rats. (G) TUNEL assays of apoptosis analysis in the rats, bar 50 μm. The experiments were replicated at least three times, mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Cell apoptosis was assessed by employing the Annexin V-FITC Apoptosis Detection Kit (CST, United States) following the manufacturer’s instruction.

Techniques: shRNA, TUNEL Assay

Journal: Frontiers in molecular biosciences

Article Title: Circular RNA TLK1 Promotes Sepsis-Associated Acute Kidney Injury by Regulating Inflammation and Oxidative Stress Through miR-106a-5p/HMGB1 Axis.

doi: 10.3389/fmolb.2021.660269

Figure Lengend Snippet: FIGURE 3 | CircTLK1 knockdown relieves LPS-induced inflammation and apoptosis in the HK-2 cells. (A) Expression of circTLK1 was detected by qPCR in HK-2 cells treated with circTLK1 shRNA. (B–E) For the induction of the cell injury model, the HK-2 cells were treated with LPS (1 mg/L) for 24 h after the cell concentration reached 80%, followed by the treatment of circTLK1 shRNA. (B) qPCR analysis of circTLK1 expression. (C) Western blot analysis of TNF-α, IL-1β, and IL-6. (D) Western blot analysis of Bax, Bcl-2, and cleaved caspase-3. (E) Flow cytometry analysis of apoptosis. The experiments were replicated at least three times, mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Cell apoptosis was assessed by employing the Annexin V-FITC Apoptosis Detection Kit (CST, United States) following the manufacturer’s instruction.

Techniques: Knockdown, Expressing, shRNA, Concentration Assay, Western Blot, Flow Cytometry

Journal: Frontiers in molecular biosciences

Article Title: Circular RNA TLK1 Promotes Sepsis-Associated Acute Kidney Injury by Regulating Inflammation and Oxidative Stress Through miR-106a-5p/HMGB1 Axis.

doi: 10.3389/fmolb.2021.660269

Figure Lengend Snippet: FIGURE 6 | CircTLK1 induces sepsis-related AKI by the miR-106a-5p/HMGB1 axis. (A–C) The CLP-induced sepsis-related AKI rat model was established in Wistar rats (N 6), followed by the indicated treatment. The cecum was exposed without perforation in the sham groups. (A) qPCR analysis of HMGB1 expression. (B) H&E staining of kidney tissues, bar 50 μm. (C) TUNEL assays of apoptosis analysis in the rats, bar 50 μm. (D–G) For the induction of the cell injury model, the HK-2 cells were treated with LPS (1 mg/L) for 24 h after the cell concentration reached 80%, followed by the indicated treatment. (D) qPCR analysis of HMGB1 expression. (E) Western blot analysis of TNF-α, IL-1β, and IL-6. (F) Western blot analysis of Bax, Bcl-2, and cleaved caspase-3. (G) Flow cytometry analysis of apoptosis, mean ± SD, **P < 0.01, ***P < 0.001.

Article Snippet: Cell apoptosis was assessed by employing the Annexin V-FITC Apoptosis Detection Kit (CST, United States) following the manufacturer’s instruction.

Techniques: Expressing, Staining, TUNEL Assay, Concentration Assay, Western Blot, Flow Cytometry