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Role of Catalase and p53 in ROS Regulation

This study explores the relationship between catalase, p53, and ROS levels in different cell lines. Results show the impact of catalase and p53 on ROS regulation.

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Role of Catalase and p53 in ROS Regulation

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  1. catalase p53 MW(kDa) 170 130 catalase 95 PIG3 MW(kDa) 72 55 72 43 55 34 43 34 26 Supplementary Figure 1 A B

  2. Vector PIG3  160   140 120 100 DCF fluorescence level 80 60 40 20 0 H1299 SW480 HCT116 p53-/- Supplementary Figure 2 A B Vector PIG3 1.4    1.2 1.0 0.8 Catalase activity (U/ml) 0.6 0.4 0.2 0.0 H1299 HCT116 p53-/- SW480 PIG3 catalase -tubulin

  3. catalase Supplementary Figure 3 U2OS RKO HCT116 siRNA Control p53 Control p53 Control p53 p53 p53R2 SESN1 SESN2 GPX1 ALDH4A1 SOD2 TIGAR Nrf-2 TP53INP1 GAPDH

  4. Supplementary Figure 4 UV (10 J/m2) H2O2 (mM) HCT116 p53-/- HCT116 HCT116 p53-/- HCT116 0 4 8 12 24 0 4 8 12 24 (hr) 0 0.05 0.1 1 2 0 0.05 0.1 1 2 p53 p53R2 catalase -tubulin UV (10 J/m2) H2O2 (mM) Saos-2 U2OS Saos-2 U2OS 0 4 8 12 24 0 4 8 12 24 (hr) 0 0.05 0.1 1 2 0 0.05 0.1 1 2 p53 p53R2 catalase -tubulin

  5. catalase p53R2 MW(kDa) 72 55 43 Supplementary Figure 5

  6.  n.s. 120  100 80 60 ROS levels (%) 40 20 0 vector wt R2 R4 Supplementary Figure 6 A B  n.s. 250  200 150 Catalase activity (%) 100 50 0 vector wt R2 R4

  7. Supplementary Figure 7 B A Control siRNA p53R2 siRNA 160 Control siRNA p53R2 siRNA  140   n.s. 120 1.8 100 1.6 ROS levels (%) 1.4  80 n.s. 1.2 60 1.0 Catalase activity (U/ml) 40 0.8 0.6 20 0.4 0 0.2 HCT116 U2OS RKO 0.0 U2OS RKO HCT116 p53R2 catalase -tubulin

  8. Supplementary Figure 8 Control vector Control vector B A Wild-type p53R2 Wild-type p53R2 Mutant p53R2 (V115L) Mutant p53R2 (V115L) n.s. n.s. n.s. 2.5   120    n.s. n.s. 2.0 100  n.s. 1.5 80 Catalase activity (U/ml) ROS levels (%) 60 1.0 40 0.5 20 0.0 U2OS RKO HCT116 0 p53R2 U2OS RKO HCT116 catalase -tubulin

  9. Supplementary Figure 9 UV (10 J/m2) UT Sup P Sup P p53 PIG3 p53R2 Catalase β-actin PMP70

  10. Supplementary Figure 10 catalase merge DAPI p53 UT p53R2 p53 UV (10 J/m2) p53R2

  11. Supplementary Figure 11 A B U2OS (vector) U2OS (vector) U2OS (catalase) U2OS (catalase) 200   35 180 160 30 140  25 120 Apoptosis (%) ROS levels (%) 20 100 80 15  60 10 40 20 5 0 0 0 8 24 h 0 24 48 h UV (40 J/m2) UV (40 J/m2)

  12. Supplementary Figure 12 H1299 (vector) 180 H1299 (catalase)  160 140 120 ROS levels (%) 100 80 60 40 20 0 Ad p53 0 48 h

  13. Supplementary Figure 13  Untreated control 200 UV (20 J/m2) 180 160 140 120 ROS levels (%) 100 80 60 40 20 0 Vector PIG3 HCT116 p53-/-

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