B A GFP-TP 53 INP 1α GFP-TP 53

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B A GFP-TP 53 INP 1α GFP-TP 53 INP 1β 6 His-Flag-SUMO 1 - + + + GFP-TP 53 INP 1βWT GFP-TP 53 INP 1βK 113 R 6 His-Flag-SUMO 1 + + + - + + k. Da 250 150 250 TP 53 INP 1 SUMO 1 (anti TP 53 INP 1) TP 53 INP 1β SUMO 1 150 100 075 * 100 SUMO 1 Ni 2+ pull down TP 53 INP 1β 100 TP 53 INP 1 100 SUMO 1 β-Tubulin SUMO 1 Input β-Tubulin Input C - 6 His-Flag- SUMO 1 Ub Nedd 8 GFP-TP 53 INP 1βWT + - + - GFP-TP 53 INP 1βK 113 R - + - + Modified TP 53 INP 1β k. Da 250 150 * 100 SUMO 1, Ub, Nedd 8 Ni 2+ pull down TP 53 INP 1β 100 SUMO 1, Ub, Nedd 8 β-Tubulin Input Supplementary Figure 1

A + - - - - + - GFP-TP 53 INP 1αWT GFP-TP 53 INP 1αK 113 R GFP-TP 53 INP 1βWT GFP-TP 53 INP 1βK 113 R 6 His-Flag-SUMO 1 + + + + k. Da 250 150 TP 53 INP 1 SUMO 1 100 075 SUMO 1 Ni 2+ pull down 100 TP 53 INP 1 075 SUMO 1 β-Tubulin Input B GFP-TP 53 INP 1αWT GFP-TP 53 INP 1αK 113 R GFP-TP 53 INP 1βWT GFP-TP 53 INP 1βK 113 R 6 His-Flag-SUMO 1 + + + + + k. Da 150 TP 53 INP 1 SUMO 1 100 075 SUMO 1 Ni 2+ pull down 100 TP 53 INP 1 075 SUMO 1 β-Tubulin Input Supplementary Figure 2

Supplementary Figure 3 U 2 OS du In le U cib 2 O S St le ab U 2 le OS St U 2 ab OS le (W U 2 T) OS (K 11 3 R ) cib du In Pon. A + - TP 53 INP 1α βTubulin

TP 53 INP 1 LC 3 DAPI Merge Untreated TP 53 INP 1αWT TP 53 INP 1αK 113 R Bafilomycin A 1 TP 53 INP 1αWT TP 53 INP 1αK 113 R Supplementary Figure 4

Supplementary Figure 5 0. 2 0 1. 2 1. 0 0. 2 0. 8 0. 6 0. 4 * 0. 8 0. 6 0. 4 0 0 1. 2 1. 0 0. 2 0 0 PIAS 4 m. RNA level si. RNA Sc Ub PIA PIA CB Ra + + + + ble P 2 n. B X 4 S 3 S 2 S 1 c 9 ram Sc 6 His-Flag-SUMO 1 TP 53 INP 1α SUMO 1 0. 2 1. 0 * 0. 8 0. 6 0. 4 * 0 Sc ram Si. R ble NA PIA S 4 0. 4 Sc r Si. R amb le NA PIA S 3 1. 2 PIAS 3 m. RNA level * Ubc 9 m. RNA level 0. 6 r Si. R amb NA le Ub c 9 * 0. 8 Sc 0. 6 BP 2 0. 8 ble 0 1. 0 Ra n 0. 4 * NA 0. 2 Sc ram Si. R ble NA PIA S 2 0. 4 ram 1. 2 PIAS 2 m. RNA level 0. 6 Ran. BP 2 m. RNA level 0. 8 Sc Sc r Si. R amb le NA PIA S 1 PIAS 1 m. RNA level B Si. R Sc ram Si. R ble NA CB X 4 CBX 4 m. RNA level A 250 150 k. Da 100 175 SUMO 1 Ni 2+ pull down TP 53 INP 1α 175 SUMO 1 Input 0. 8 0. 6 0. 4 0. 2 *

Supplementary Table 1 Targeted gene Si. RNA sequence Scramble 5’-UUCUCCGAACGUGUCACGU-3’ PIAS 1 5’-AAUCCGGAUCAUUCUAGAGCU-3’ PIAS 2 5’-ACAGAAGCGCCCUGGACGC-3’ PIAS 3 5’-GGAGCCAAAUGUGAUUAUA-3’ PIAS 4 5’-GUACUUAAACGGACUGGGA-3’ Ran. BP 2 5’-GAAUAACUAUCACAGAAUG-3’ CBX 4 5’-UCGCCUUCCAGAACAGGGAAC-3’ Ubc 9/UBE 2 I 5’-UAAAUUCGAACCACCAUUA-3’ SENP 1 5'-AACUACAUCUUCGUGUACCUC-3' SENP 2 5'-GGUAACUUCUGCUUGUAAU-3’ SENP 6 5'-GAAAGUGACCCUCGUUAUA-3' Supplementary Table 2 Gene Forward primer Reverse primer p 21 5’-CCTGTCACTGTCTTGTACCCT-3’ 5’-GCGTTTGGAGTGGTAGAAATCT-3’ Bax 5’-CCCCGAGAGGTCTTTTTCCG-3’ 5’-GGCGTCCCAAAGTAGGAGA-3’ PUMA 5’-GACCTCAACGCACAGTACGAG-3’ 5’-AGGAGTCCCATGATGAGATTGT-3’ PIAS 1 5’-ACTGGAACTCCCACATCTTACA-3’ 5’-GTTGTCTGATGCTAGACTGGTG-3’ PIAS 2 5’-GGCTTTGCTGGACGGAATAAA-3’ 5’-CGGCGTCTATACAATTCTCGGAT-3’ PIAS 3 5’-GGTTTGAGGAAGCGCACTTTA-3’ 5’-CTCCTGGCAGAACCTCTCT-3’ PIAS 4 5’-GTGGGCCGGAGTAAGAGTG-3’ 5’-TCAGGGCTACAGTCAAACTGC-3’ Ran. BP 2 5’-AAACCTCCGATTGCAGCTCAT-3’ 5’-GGCAAAGATGGCCTTAATCCT-3’ CBX 4 5’-AAGAACGGACGCATC-3’ 5’-CTCGCCGGACTTGATCTTCA-3’ Ubc 9/UBE 2 I 5’-AAAAATCCCGATGGCACGATG-3’ 5’-CTTCCCACGGAGTCCCTTTC-3’ Cyclophilin (house keeping gene) 5’-ATGGTCAACCCCACCGTGT-3’ 5’-TTCTTGCTGTCTTTGGAACTTTGTC-3’

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