Fig. 4: The combination treatment exerts its effects through DKK1-Mediated Ferroptosis.

A qRT-PCR and Western blot were used to detect the mRNA and protein expression of DKK1, respectively, to assess the overexpression efficiency in SK-OV-3 and A2780 cells. The qRT-PCR was used GAPDH as the reference gene and analyzed using the 2^-ΔΔCT method relative to control, based on three biological replicates. Data are presented as means ± SD and were analyzed using Student’s t test. B The DCFH-DA fluorescent probe was used to detect intracellular ROS in cells treated for 48 h under different conditions, accompanied by a bar plot depicting relative intracellular ROS expression. Data are presented as means ± SD from three biological replicates and were analyzed using Student’s t test. C Flow cytometry with Annexin V-FITC/PI double staining was used to detect apoptosis levels in control cells and DKK1-overexpressing cells (DKK1 OE) after 48 h of different treatments, accompanied by a bar plot depicting the proportion of dead cells. Data are presented as means ± SD from three biological replicates and were analyzed using Student’s t test. D The MitoSOX fluorescent probe was used to detect mitochondrial ROS in cells after 48 h of different treatments, accompanied by a bar plot depicting relative mitochondrial ROS expression. Data are presented as means ± SD from three biological replicates and were analyzed using Student’s t test. E Western blot was used to detect DKK1 protein expression in control cells and DKK1-overexpressing cells (DKK1 OE) after 48 h of different treatments. Experimental groups are defined as follows: 1 represents the Control group; 2 represents the KD01 + Erastin combination treatment group; 3 represents the DKK1 overexpression Control group; and 4 represents the DKK1 overexpression combined with KD01 + Erastin treatment group. Statistical significance was determined as follows: *p < 0.05, **p < 0.01, ***p < 0.001, NS > 0.05.