Figure Legends:
Figure 1. The top 50 signaling pathways regulated by Bazhen decoction.
BZT C1-C3 were three control samples, BZT L1-L3 were three samples with low dose (10μg/mL) Bazhen decoction treatment, BZT H1-H3 were three samples with high dose (100μg/mL) Bazhen decoction treatment. The red color stands for up-regulation, the blue color stands for down-regulation.
Figure 2. The Bazhen decoction activated the DNA helicase and telomere maintenance pathways. A. The DNA helicase and telomere maintenance pathways were up-regulated by Bazhen decoction. B-C. After Bazhen decoction treatment, the protein level of DNA helicase Blm, Mcm7, Parp1 and telomere protein Terf1were up-regulated, while telomere protein Pot1 was down-regulated in WT cells (B) or G3DKO cells (C).
D-E. After Bazhen decoction treatment, the protein level of cell cycle inhibitor p21, Rb, and p130 were up-regulated, while cell cycle promoter E2F1 was also up-regulated in WT cells (D) or G3DKO cells (E).
Figure 3. In G3DKO cells, the Bazhen decoction up-regulated the DNA helicase, promoted the resolving of G4 structure.
The signals of G4 structure were diminished by Bazhen decoction, while the Recql4 signals were elevated. The PDS treatment stabilized the G4 signals, and down-regulated the Recql4, while combined Bazhen decoction rescued the Recql4 expression and diminished the G4 signals.
The signals of G4 structure were diminished by Bazhen decoction, while the Blm signals were elevated. The PDS treatment stabilized the G4 signals, and down-regulated the Blm, while combined Bazhen decoction rescued the Blm expression and diminished the G4 signals.
Figure 4. In G3DKO cells, the Bazhen decoction regulated cell cycle progression, and protected the cells from DNA damages induced by G4 stabilizer PDS.
  1. In G3DKO cells, PDS treatment down-regulate the DNA helicase Blm, Recql4, Parp1, Mcm7, telomere protein Terf1, and cell cycle protein Cdc2, Cdk2, while Bazhen decoction rescued the expression of these proteins.
  2. The flow cytometry analysis of G3DKO cells showed that the percentage of G1 phase cells was reduced and the percentage of S phase and G2 phase was increased. The PDS treatment also induced the increase of G2 cells.
  3. The Bazhen decoction application reduced the signal of DNA damage marker γ-H2AX in the G3DKO cells. The cells treated with PDS showed increasedγ-H2AX signals, while the application of Bazhen decoction obviously diminished theγ-H2AX signals induced by PDS.
Figure 5. Bazhen decoction treatment facilitate the elongation of telomere.
  1. Real-time PCR data showed that the telomere DNA increased significantly after Bazhen decoction treatment (p<0.01).
  2. IF-FISH showed that the DNA helicase Recql4 was up-regulated (Recql4), and the telomere DNA signals significantly increased after Bazhen decoction treatment (Telomere), the colocalization of Recql4 with telomere DNA increased (Telomere & Recql4, white arrows pointed to the colocalization spots).
  3. IF-FISH showed that the DNA helicase Blm was up-regulated (Blm), and the telomere DNA signals significantly increased after Bazhen decoction treatment (Telomere), the colocalization of Blm with telomere DNA increased (Telomere & Blm, white arrows pointed to the colocalization spots).