Phospho-Histone H2A.X (Tyr143) Antibody - #AF8482

Figure 3. Shikonin inhibits PLY-induced nuclear DNA damage. (A) The solvent or shikonin solution does not lead to DSBs in cells. (B) Nuclear γH2AX foci in cells exposed in vitro to rPLY for 6 hours. (C) Histogram of DSBs foci, ** indicates p <0.01 compared with the shikonin-free group.

Figure 4 A combination of Chidamide and Bortezomib increases production of ROS dependent DNA damage and the changes of cell apoptosis and cycle pathway in MM cells. (A) ARP-1 cells were pretreated with or without NAC (15.0 mmol/L) for 2 hours at 37°C and then incubated with CHI (1.0 µmol/L) and/or BTZ (5.0 nmol/L) for 24 hours, then ROS generation was detected. (B) ARP-1 cells were pretreated with or without 15 mmol/L NAC and then treated with Chidamide or Bortezomib alone or in combination, and cell viabilities were evaluated using CCK-8 assays. (C) The expression of γ-H2AX in ARP-1 cells treated with CHI (1.0 µmol/L) and/or BTZ (5.0 nmol/L) were determined by Western blot. (D) Representative images of γ-H2AX (Red) and nuclei (Blue) in ARP-1 cells treated with single agent or combination for 24 hours by immunofluorescence assay. Scale bars represent 20 µm. (E, F) Western blot analysis of the expressions of cleaved caspase3, cleaved caspase8, cleaved PARP-1 and HDAC1 in XG1 (E) and ARP-1 (F) cells after 48 hours treatment with single agent or in combination. Error bars indicate mean ± SD. **p < 0.01.

Figure 4 A combination of Chidamide and Bortezomib increases production of ROS dependent DNA damage and the changes of cell apoptosis and cycle pathway in MM cells. (A) ARP-1 cells were pretreated with or without NAC (15.0 mmol/L) for 2 hours at 37°C and then incubated with CHI (1.0 µmol/L) and/or BTZ (5.0 nmol/L) for 24 hours, then ROS generation was detected. (B) ARP-1 cells were pretreated with or without 15 mmol/L NAC and then treated with Chidamide or Bortezomib alone or in combination, and cell viabilities were evaluated using CCK-8 assays. (C) The expression of γ-H2AX in ARP-1 cells treated with CHI (1.0 µmol/L) and/or BTZ (5.0 nmol/L) were determined by Western blot. (D) Representative images of γ-H2AX (Red) and nuclei (Blue) in ARP-1 cells treated with single agent or combination for 24 hours by immunofluorescence assay. Scale bars represent 20 µm. (E, F) Western blot analysis of the expressions of cleaved caspase3, cleaved caspase8, cleaved PARP-1 and HDAC1 in XG1 (E) and ARP-1 (F) cells after 48 hours treatment with single agent or in combination. Error bars indicate mean ± SD. **p < 0.01.

Fig 4. |Betulin inhibits PLY-induced DNA damage. The A549 cells were exposed in vitro to PLY with or without betulin for 6 h. (A) The images were selected from immunofluorescence assays and were captured by confocal microscopy.

Figure 6 Combination of chidamide (CHI) and lenalidomide (Len) increase ROS-related DNA damage in MM cells. (A and B) DNA damage analysis by γ-H2AX foci immunofluorescence. Representative images of γ-H2AX (Red) and nuclei (Blue) in ARP-1 and RPMI-8226 cells treated with single agents or combination for 24 hours. Scale bars represent 20 μm. (C and D) after treated with 1 μM chidamide and/ or 4 μM lenalidomide for 24 hours, expression of γ-H2AX was determined using Western blotting.

Figure 6 Combination of chidamide (CHI) and lenalidomide (Len) increase ROS-related DNA damage in MM cells. (A and B) DNA damage analysis by γ-H2AX foci immunofluorescence. Representative images of γ-H2AX (Red) and nuclei (Blue) in ARP-1 and RPMI-8226 cells treated with single agents or combination for 24 hours. Scale bars represent 20 μm. (C and D) after treated with 1 μM chidamide and/ or 4 μM lenalidomide for 24 hours, expression of γ-H2AX was determined using Western blotting.