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FIGURE 1 A p38 EV Sh MAPK 14 HCT116-TP53KO GAPDH EV Sh MAPK 14bis Sh MAPK 12 Sh MAPK 13 EV Sh MAPK 11 B 0.0 0.5 1.0 1.5 2.0 2.5 Sh LUC Sh MAPK 14 Sh MAPK.

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Presentation on theme: "FIGURE 1 A p38 EV Sh MAPK 14 HCT116-TP53KO GAPDH EV Sh MAPK 14bis Sh MAPK 12 Sh MAPK 13 EV Sh MAPK 11 B 0.0 0.5 1.0 1.5 2.0 2.5 Sh LUC Sh MAPK 14 Sh MAPK."— Presentation transcript:

1 FIGURE 1 A p38 EV Sh MAPK 14 HCT116-TP53KO GAPDH EV Sh MAPK 14bis Sh MAPK 12 Sh MAPK 13 EV Sh MAPK 11 B Sh LUC Sh MAPK 14 Sh MAPK 14bis Sh MAPK 11 Sh MAPK 12 Sh MAPK 13 IC50 (nM) HCT116-TP53KO HA MAPK14 CA MAPK11 CAMAPK12 CA MAPK13CA tubulin HCT116-TP53KO D Kinase Assay EV MAPK 14 CA MAPK 11 CA MAPK 12 CA MAPK 13 CA IC50 (nM) HCT116-TP53KO E pp38 GAPDH HCT116-TP53KO NTSN38 F C p ATF2 MAPK14 CA MAPK11 CAMAPK12 CAMAPK13CA

2 FIGURE 2 A HCT116-TP53KO EV HCT116-TP53KO-MAPK14CA Cell number (10 6 ) Days B Doubling Time (hours) EVMAPK14-CA HCT116-TP53KO HCT116-TP53KO-EV HCT116-TP53KO-MAPK14CA D7D14D21D28 % living cells C HCT116-TP53 KO EV MAPK14 CA % APOPTOSIS D E HCT116-TP53KO-EVHCT116-TP53KO-MAPK14CA HCT116- TP53KO -EV HCT116- TP53KO- MAPK14CA Blue cell number/field

3 FIGURE 3 A HCT116-TP53KO-EVHCT116-TP53KO-MAPK14CA S 49.4% G0 G1 28.8% G2/M 18.4% S 37.1% G0 G1 36.8% G2/M 19.3% B HCT116-TP53KO-EV HCT116-TP53KO-MAPK14CA % of BrdU stained cells D EVD7D14D21D28 p21 Arbitrary RNA units MAPK14CA HCT116-TP53KO C p21 Actin EVD7D14D21D28 MAPK14CA HCT116-TP53KO HA p21 shLUC shp21 HCT116-TP53KO-MAPK14CA GAPDH E HCT116-TP53KO -ShLuc-MAPK14CA HCT116-TP53KO -Shp21-MAPK14CA S 25.8% G0 G1 45.6% G2/M 23.16% S 46.1% G0 G1 20.9% G2/M 28.9% F G HCT116-TP53KO-ShLuc- MAPK14CA HCT116-TP53KO-Shp21- MAPK14CA % of BrdU stained cells

4 FIGURE 4 A HCT116-TP53KO EVMAPK14CA LC3 Actin EVD7D14D21D28 MAPK14CA HCT116-TP53KO H1299 LC3 Tubulin EVD7 D14 D21 MAPK14CA B C 500 nm HCT116-TP53KO EVMAPK14CA Autophagosome number/cell MAPK14CAEV HCT116-TP53 KO D MAPK14CAEV GFP-LC3 labelling E HCT116-TP53KO- MAPK14CA GAPDH LC3 shLUC shp21 HCT116-TP53KO GFP-LC3 spots/cell EV MAPK14 CA

5 FIGURE 5 p62 Actin EVD7D14D21D28 MAPK14CA HCT116-TP53KO A EVD7D14D21D28 p62 level (arbitrary unit) MAPK14CA HCT116-TP53KO B HCT116-TP53KO (J14) p62 NT2H4H8H NT2H4H8H EVMAPK14CA GAPDH nt2h4h8hnt2h4h8h p62 level (arbitrary unit) EV HCT116-TP53KO MAPK14CA NT 3MA Baf NT Baf 3MA LC3 Actin EV HCT116-TP53KO MAPK14CA D14 HCT116-TP53KO EV MAPK14CA D NT3MABafNT3MABaf LC3-II level (arbitrary unit) C D E Atg7 Actin HCT116-TP53KO Atg5 siCT D2 D4 siRNA NT 3MA Baf HCT116-TP53KO EV MAPK14CA D14 Cell viability (%) HCT116-TP53KO EV MAPK14CA D14 Cell viability (%) siCT siAtg 5siAtg 7 siCT siAtg 5siAtg 7

6 FIGURE 6 A LC3 labelling HCT116-TP53 KO SN38 NT LC3 spots/cell NTSN38 HCT116-TP53KO B NTSN HCT116 TP53KO LC3 Actin NTSN SW480 LC3 Actin C SN38 SN+ Baf SN+ 3MA LC3 GAPDH HCT116-TP53KO D siCT siAtg5 siAtg7 SW480 SN38 IC50 (nM) Cytotoxic Assay EF siCT siAtg5 siAtg HCT116-TP53KO SN38 IC50 (nM) Cytotoxic Assay siCTsiATG5siATG7 HCT116-TP53KO SN38 IC50 (nM) Clonogenic Assay HCT116-TP53KO NTSN38SN38+3MA % living cells G

7 FIGURE 7 LC3 MAPK14 Actin NT SN SN+3MA HCT116-TP53KO- shLUC HCT116-TP53KO- shMAPK14 A B shLUCshMAPK 11 shMAPK 12 shMAPK 13 shLUCshMAPK 11 shMAPK 12 shMAPK 13 LC3 HCT116-TP53KO GAPDH NTSN HCT116-TP53KO ShLuc HCT116-TP53KO ShMAPK14 SN38 SN38+3MA SN38 IC50 (nM) C D siCT siATG5 siATG HCT116-TP53KO ShLuc SN38 IC50 (nM) HCT116-TP53KO ShMAPK14


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