SP2 Antibody - #DF8721

Figure 4 Transcription of the Dapk3 gene is promoted by Sp2. A) Three potential REs of the Dapk3 gene were predicted by the JASPAR database. B–G) TM3 cells were transfected with 0, 2, and 4 µg pcDNA3.1-Sp2 for 48 h or 250 pmol µL−1 si-Sp2 for 24 h, and the mRNA and protein levels of DAPK3 were determined. H–J) After TM3 cells were exposed to 0 or 400 µm DBP for 24 h in the presence or absence of 250 pmol µL−1 si-Sp2, the mRNA and protein contents of DAPK3 were detected. K) After TM3 cells were transfected with pcDNA3.1 or pcDNA3.1-Sp2 in the presence or absence of pGL4.2-Dapk3 prom, the luciferase activity was detected. L) TM3 cells were transfected with pGL4.2-RE1, pGL4.2-RE2, or pGL4.2-RE3 together with or without pcDNA3.1-Sp2, the luciferase activity was then determined. M) After TM3 cells were transfected with pcDNA3.1 or pcDNA3.1-Sp2 in the presence of pGL4.2-RE3 or pGL4.2-mutRE3, the luciferase activity was determined. N) The binding of Sp2 to the RE3 site of the Dapk3 gene was verified by ChIP-qPCR. Data are represented as means ± SEM, n = 3. *P < 0.05, by one-way ANOVA with LSD method (B, D, H, J, K, and M) or independent sample t-test (E, G, L, and N).

Fig. 2. Sp2 is essential for DBP-induced ferroptosis of mouse Leydig cells. (A) Adult male mice were exposed to 0–500 mg/kg DBP for 28 d, and the expression of Sp2 in the testis tissue was determined by immunohistochemistry. (B, C) The mRNA and protein levels of Sp2 were detected after TM3 cells were treated with 0–400 μM DBP for 24 h. The protein levels of TFR and GPX4 (D), the mitochondrial morphology (E), and the intracellular Fe2+ concentration (F, G) were determined after TM3 cells were transfected with 0, 2 and 4 μg pcDNA3.1-Sp2 for 48 h. The protein levels of TFR and GPX4 (H), and the intracellular Fe2+ concentration (I, J) were detected after TM3 cells were transfected with 250 pmol/μl si-Sp2 for 24 h. The protein levels of TFR and GPX4 (K), the intracellular Fe2+ concentration (L, M), and the mitochondrial morphology (N) were determined after TM3 cells were treated with 0 or 400 μM DBP for 24 h with or without Sp2 knockdown. *P < 0.05.