Iκbα 44d4 4812

Temozolomide (34219) was obtained from Sigma‐Aldrich. SC75741 (HY‐10496), SN50 (HY‐P0151), JSH‐23 (HY‐13982), and lomeguatrib (HY‐13668) were obtained from MedChemExpress. Primary antibodies against NF‐κB p65 (D14E12) (59674S, 1:1000), p‐NF‐κB p65 (Ser536) (93H1) (3033S, 1:1000), and IκBα (44D4) (4812, 1:1000) were purchased from Cell Signaling Technology. Primary antibodies against MGMT (ab69629) were purchased from Abcam. Primary antibodies against Lamin B1 (B‐10) (sc‐374015) were purchased from Santa Cruz Biotechnology. Primary antibodies against GAPDH (1A6) (MB001, 1:1000) were purchased from Bio‐World. Anti‐mouse IgG, HRP‐linked antibody, and anti‐rabbit IgG, HRP‐linked antibodies were purchased from Santa Cruz Biotechnology. Matrigel (354248) was obtained from BD Biosciences.

For transient expression in mammalian cells, full open reading frames (ORF) for TRIM1 and HIF1α were amplified using a SW480 cDNA library and inserted into the pCS2-EGFP and pCS2-Flag vectors. pRK5-HA-Ub-WT and the lysine mutants plasmids were maintained in our lab [35 (link)]. HRE-luc, pNF-κB-Luc, and pRL-TK reporter plasmids were purchased from Addgene. The sequences of all plasmids were confirmed by sequencing before use.
Antibodies for GAPDH (G9545) and Flag (F7425) were purchased from Sigma-Aldrich. EGFP (sc8334) antibody was obtained from Santa Cruz Biotechnology. Antibodies for HIF1α antibodies (D1S7W, #36169S), NF-κB p65 (D14E12, #8242), phospho-NF-κB p65 (Ser536) (93H1, #3033) and IκBα (44D4, #4812) were from Cell Signaling Technology. Anti-TRIM1/MID2 (68359-1-Ig) and anti-HA Epitope Tag (901501) antibodies were from Proteintech and Biolegend. DMOG was from Selleckchem. Cell culture products were from Invitrogen. The relevant chemicals in this study were obtained from Sigma-Aldrich unless stated otherwise.