Skip to main content
. 2021 May 7;24(6):102521. doi: 10.1016/j.isci.2021.102521

Figure 5.

Figure 5

CAN inhibited the mTOR-HIF-1 signaling pathway in vivo and in vitro

(A–H) HIF-1a, p-mTOR (Ser 2448), and mTOR expression levels in (A) the hearts of the HFD/STZ-induced diabetic mice and in (B) the HL-1 cells were determined using western blot analysis. Relative expression level of (C) Hif-1a mRNA and quantification of (D) HIF-1α protein in the hearts of the HFD/STZ-induced diabetic mice was done using ImageJ software following western blot analysis. Relative expression levels of (E) p-mTOR (Ser 2448)/mTOR protein expression ratio in the hearts of the HFD/STZ-induced diabetic mice was determined using ImageJ following western blot analysis. Relative expression level of (F) Hif-1a mRNA and the quantification of (G) HIF-1α protein in the HL-1 cells was done using RT-qPCR and ImageJ software following western blot analysis. Relative expression levels of (H) p-mTOR (Ser 2448)/mTOR protein expression ratio in the HL-1 cells was determined using ImageJ software following western blot analysis (H).

(I) Immunofluorescence analysis of the HL-1 cells using a confocal microscope (I). Magnification, ×600. Green fluorescence indicates p-mTOR (Ser 2448), red fluorescence indicates HIF-1α, blue fluorescence indicates DAPI, and merged indicates merged signal of p-mTOR (Ser 2448), HIF-1α, and DAPI. Data are presented as the mean ± SD. n = 5 or 3. #p < 0.05 and ##p < 0.01 versus normal or blank control; ∗p < 0.05 and ∗∗p < 0.01 versus HFD/STZ or PA. HFD, high-fat diet; STZ, streptozotocin; CAN, canagliflozin; normal, normal control group; HFD/STZ, HFD/STZ-induced diabetic control group; HFD/STZ + CAN, canagliflozin-treated HFD/STZ-induced diabetic group; PA, palmitic acid; Ctrl, untreated normal or blank control; Mod/PA, PA (0.25 mM)-treated control group; 0.625, HL-1 cells were treated with PA (0.25 mM) and CAN (0.625 μg/mL); 1.25, HL-1 cells were treated with PA (0.25 mM) and CAN (1.25 μg/mL); 2.5, HL-1 cells were treated with PA (0.25 mM) and CAN (2.5 μg/mL); PA + CAN, HL-1 cells treated with PA (0.25 mM) and CAN (2.5 μg/mL); p, phosphorylated.