Anti-antioxidant impacts of circZNF609 silence in HaCaT cells through regulating miR-145

Figures & data

Figure 1. H₂O₂ evoked oxidative stress damage in HaCaT cells. (A) HaCaT cells were stimulated with various doses of H₂O₂ for 12 h. The viability of HaCaT cells was examined by CCK-8. The following experiments applied 300 μM H₂O₂ in HaCaT cells. (B) Expression of p53 and p16, (C) apoptosis rate, (D) expression of Bax, Cleaved-Caspase-3 and Cleaved-PARP, as well as (E) ROS level were examined by Western blot, FITC-PI double staining, and DCFH-DA probe. ** and *** indicate p-values less than .01 and .001.

Figure 2. cZNF609 expression induced by H₂O₂ in HaCaT cells. HaCaT cells were stimulated with 300 μM H₂O₂ for 12 h. cZNF609 level was checked by qRT-PCR. ** indicates p-value less than .01.

Figure 3. cZNF609 silence alleviated H₂O₂-evoked oxidative stress damage in HaCaT cells. (A) si-NC or si-cZNF609 was transfected into HaCaT cells. cZNF609 level was checked by qRT-PCR. Following transfection, the cells were stimulated with 300 μM H₂O₂ for 12 h. (B) Cell viability, (C,D) expression of p53 and p16, (E) apoptosis rate, (F,G) expression of Bax, Cleaved-Caspase-3 and Cleaved-PARP, as well as (H) ROS level were examined by CCK-8, Western blot, FITC-PI double staining, and DCFH-DA probe. *, ** and *** indicate p-values less than .05, .01 and .001.

Figure 4. cZNF609 silence induced miR-145 up-regulation. Following transfection with si-NC or si-cZNF609, HaCaT cells were stimulated with 300 μM H₂O₂ for 12 h. miR-145 level was checked by qRT-PCR. * and *** indicate p-values less than .05 and .001.

Figure 5. cZNF609 silence alleviated oxidative stress damage through miR-145. (A) NC or miR-145 inhibitor was transfected into HaCaT cells. miR-145 level was checked by qRT-PCR. The cells were transfected with si-cZNF609 alone or together with miR-145 inhibitor. si-NC and NC were transfected as negative controls. The transfected cells were then stimulated with 300 μM H₂O₂ for 12 h. (B) Cell viability, (C,D) expression of p53 and p16, (E) apoptosis rate, (F,G) expression of Bax, Cleaved-Caspase-3 and Cleaved-PARP, as well as (H) ROS level were examined by CCK-8, Western blot, FITC-PI double staining, and DCFH-DA probe. *, ** and *** indicate p-values less than .05, .01 and .001.

Figure 6. The decrease of miR-145 promoted cZNF609 expression. The cells were transfected with si-cZNF609 alone or together with miR-145 inhibitor. si-NC and NC were transfected as negative controls. The transfected cells were then stimulated with 300 μM H₂O₂ for 12 h. (A) cZNF609 and (B) miR-145 expression was tested by qRT-PCR. *, ** and *** indicate p-values less than .05, .01 and .001.

Figure 7. cZNF609 silence inhibited JNK and p38MAPK pathways through miR-145. HaCaT cells were transfected with si-cZNF609 alone or together with miR-145 inhibitor. si-NC and NC were transfected as negative controls. The transfected cells were then stimulated with 300 μM H₂O₂ for 12 h. The activation of (A,B) JNK and (C,D) p38MAPK was checked by Western blot. *, ** and *** indicate p-values less than .05, .01 and .001.