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.
- 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.
- 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.
- 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.
- Real-time PCR data showed that the telomere DNA increased
significantly after Bazhen decoction treatment (p<0.01).
- 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).
- 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).