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

Figure 3.

Figure 3

The protective effects of CAN on PA-induced HL-1 cell

(A) Oil red O staining results of the HL-1 cell line and quantitative analysis. Magnification, ×400.

(B) ROS was detected using the fluorescent probe, 2′,7′-dichlorofluorescein diacetate and quantitative analysis. Magnification, ×400.

(C) WGA staining results of the HL-1 cells after different treatments.

(D) Cell viability in the HL-1 cells was detected using MTT assay.

(E–H) (E) LDH activity in the HL-1 cell culture medium. mRNA expression levels of the inflammatory factors, (F) Il-1α, (G) Il-6, and (H) Tnf-α in the HL-1 cells determined using reverse transcriptase-quantitative PCR.

(I) Immunofluorescence staining with YF488-Annexin V and PI in the HL-1 cells, using a confocal microscope (magnification, ×600). Red fluorescence indicates PI, green fluorescence indicates YF488-annexin V, and the merged figure indicates the merged signal of PI and YF488-annexin V.

(J) Flow cytometry assay in the HL-1 cells. Data are presented as mean ± SD (n = 3). #p < 0.05 and ##p < 0.01 versus ctrl; ∗p < 0.05 and ∗∗p < 0.01 versus PA. Ctrl, untreated normal control; Mod/PA, PA-treated (0.25 mM) 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); ROS, reactive oxygen species, LDH, lactate dehydrogenase; PA, palmitic acid; CAN, canagliflozin.