REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
GAPDH	HuaAn Biotechnology	M1310-2
FLAG	Sigma-Aldrich	F7425
HA	Cell Signaling Technology	3724S
UFM1	Abcam	Ab109305
Anti-Flag M2 Affinity Gel	Sigma-Aldrich	A2220
Goat polyclonal anti-mouse-HRP	Sigma-Aldrich	A9169
Goat polyclonal anti-rabbit-HRP	Sigma-Aldrich	A9044
Anti-p53	Santa Cruz	sc-126
Bacterial and virus strains
DH5α	TIANGEN	CB101
BL21 (DE3)	TIANGEN	CB105
Chemicals, peptides, and recombinant proteins
Dulbecco’s Modified Eagle’s Medium(DMEM)	Biological Industries	06-1055-57-1ACS
Fetal Bovine Serum (FBS)	Biological Industries	04-001-1ACS
Trypsin EDTA solution A (0.25%)	Biological Industries	03-050-1A
Opti-MEM™ I Reduced Serum Medium(Opti-MEM)	Gibco	31985070
Penicillin-Streptomycin	Biological Industries	03-031-1B
Polyethyleneimine (PEI)	Polysciences	23966
N-Ethylmaleimide (NEM)	Sigma-Aldrich	E3876
Imidazole	Biosharp	1-1025
NP40 Alternative	Sigma-Aldrich	492016
cOmplete Protease Inhibitor Cocktail Tablets	Roche	11697498001
Sodium Chloride	Sigma-Aldrich	7647-14-5
PMSF	Roche	329-98-6
SDS	Sigma-Aldrich	151-21-3
MgCl2	Sigma-Aldrich	7786-30-3
Recombinant Human His6-UFM1 Activating Enzyme (UBA5)	R&D Systems	E-319-025
Recombinant Human His6-UFC1 Protein (UFC1)	R&D Systems	E2-675-100
Recombinant Human His6-UFM1 Protein (UFM1)	R&D Systems	UL-500-250
p53 Protein	R&D Systems	SP-454-020
Recombinant Human UFL1	Abnova	H00023376-P01
Human DDRGK1 protein (Recombinant Myc-DDK (FLAG)) (Full Length)	LSBio	LS-G72211-20
IPTG	Sigma-Aldrich	16758
Critical commercial assays
SuperSignalTM West Femto Maximum Sensitivity Substrate	Thermo Fisher Scientific	34095
ATP (10 mM)	Cell Signaling Technology	9804S
PreScission protease	Beyotime	P2303
Experimental models: Cell lines
HEK293T cells	ATCC	CRL-11268
Recombinant DNA
pSG5-HA	Liu et al. (2020)	N/A
pSG5-HA-UFM1	Liu et al. (2020)	N/A
pSG5-HA-UFM1-ΔC2	Liu et al. (2020)	N/A
pSG5-HA-UFM1-ΔC3	Liu et al. (2020)	N/A
pSG5-HA-UBA5	Liu et al. (2020)	N/A
pSG5-HA-UFC1	Liu et al. (2020)	N/A
pSG5-HA-UFL1	Liu et al. (2020)	N/A
pSG5-HA-DDRGK1	Liu et al. (2020)	N/A
pCDNA3.0-FLAG-His	Liu et al. (2020)	N/A
pCDNA3.0-FLAG-His-p53	Liu et al. (2020)	N/A
pGEX-6P-1	Liu et al. (2020)	N/A
pGEX-6P-1-UFL1	Liu et al. (2020)	N/A
pGEX-6P-1-DDRGK1	Liu et al. (2020)	N/A
Software and algorithms
Image Lab	Bio-Rad Laboratories	https://www.bio-ad.com
Other
Heraeus Multifuge X3R	Thermo Fisher Scientific	41270886
Centrifuge	Eppendorf	5424R
Centrifuge	Eppendorf	5424
ChemiDocTM MP Imaging System	Bio-Rad Laboratories	734BR4093
Ultrasonic Cell Disruptor	Tengxiao	TX650