Fig. 2: SPOP is translocated to the cytoplasm upon LPS stimulation. | Cellular & Molecular Immunology

Fig. 2: SPOP is translocated to the cytoplasm upon LPS stimulation.

From: SPOP negatively regulates Toll-like receptor-induced inflammation by disrupting MyD88 self-association

Fig. 2

(a) SPOP was expressed both in the cytoplasm and the nucleus. Cytoplasmic and nuclear fractions from THP-1 cells (3 × 107) were isolated, equilibrated to equal volumes and analyzed by immunoblots with the indicated antibodies. (b) Cell fractionation and immunoblot analysis of the subcellular fractions. BMDMs (3 × 107) were treated with LPS (100 ng/mL) for the indicated times. Cell fractionations were performed; the fractions were equilibrated to equal volumes and analyzed by immunoblots with the indicated antibodies (Upper). The whole cellular levels of SPOP upon LPS stimulation were analyzed by immunoblot with anti-SPOP (Lower). (c) Confocal microscopy of the cellular localization of SPOP. HEK293 cells (1 × 105) were transfected with expression plasmids for TLR4-Flag (100 ng), MD2-Flag (50 ng) and mCherry-SPOP (50 ng). Twenty hours after transfection, cells were treated with LPS (100 ng/mL) or left untreated for 1 hour, and then fixed with 4% paraformaldehyde and subjected for confocal microscopy. Cyt, cytoplasm; Nuc, nucleus.

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