Journal: Scientific Reports

Article Title: TRIB1 confers therapeutic resistance in GBM cells by activating the ERK and Akt pathways

doi: 10.1038/s41598-023-32983-w

Figure Lengend Snippet: TRIB1 protein level oscillates during cell cycle and TRIB1 may cause p53 degradation through COP1. ( a ) Protocol schematic for double thymidine block in U87 MG cells. ( b ) Western blot depicting protein levels of TRIB1 and respective cell cycle proteins at indicated time points following thymidine release. ( c ) Protocol schematic for G2 arrest by Ro-3306 in U87 MG cells. ( d ) Western blot for protein levels of TRIB1 and respective cell cycle proteins 30 min after release from Ro-3306 treatment. Western blot showing the co-immunoprecipitation of TRIB1 with p53, COP1 and HDAC1 in ( e ) T08-387-TRIB1-FLAG cells and ( f ) GBM30-TRIB1-FLAG cells (n = 3). ( g ) Western blot showing the coimmunoprecipitation of TRIB1 with p53, COP1 and HDAC1 in 3359 parental and 3359-TP53 R248W cells after TRIB1 overexpression (n = 3). ( h ) Western blot showing co-immunoprecipitation of p53 and COP1 with TRIB1 in T08-387 cells after COP1 knockdown by shRNA. n = 2 for all western blots unless otherwise mentioned. For cell cycle experiment ( b , d ), depending on the molecular weight, one or more proteins were probed on same blot after cutting. For immunoprecipitation, each protein was probed on separate blots.

Article Snippet: Control shRNA plasmid-A (sc-108060) and COP1 shRNA Plasmid (h) (sc-45541-SH) were purchased from Santa Cruz biotechnologies.

Techniques: Blocking Assay, Western Blot, Immunoprecipitation, Over Expression, Knockdown, shRNA, Molecular Weight