Fig. 5: FLOT1/2 are co-purifying with M1 poly-Ub during necroptosis progression.
From: LUBAC-mediated M1 Ub regulates necroptosis by segregating the cellular distribution of active MLKL
A UBAN-mediated pulldown of M1 ubiquitinated proteins in control or HOIPIN-8 (30 µM)-pre-treated HT-29 cells treated with TBZ (10 ng/mL TNFα, 1 µM BV6, 20 µM zVAD.fmk) for the indicated time points. GST and GAPDH were used as loading controls. Representative blots of at least two independent experiments are shown. B UBAN-mediated pulldown of M1 ubiquitinated proteins in control, HOIPIN-8 (30 µM), Nec-1s (30 µM), GSK‘872 (20 µM) or NSA (10 µM)-pre-treated HT-29 cells treated with TBZ (10 ng/mL TNFα, 1 µM BV6, 20 µM zVAD.fmk) for 4 h. GST and GAPDH were used as loading controls. Representative blots of at least two independent experiments are shown. C GST-UBAN-mediated pulldown of M1 ubiquitinated proteins in control, HOIPIN-8 (30 µM) or NSA (10 µM)-pre-treated HT-29 cells treated with TBZ (10 ng/mL TNFα, 1 µM BV6, 20 µM zVAD.fmk) for 4 h. GST and GAPDH were used as loading controls. Representative blots of at least two independent experiments are shown. See also Figure S5.
