Figure 3 | Cell Death & Disease

Figure 3

From: Ubiquitylation of MFHAS1 by the ubiquitin ligase praja2 promotes M1 macrophage polarization by activating JNK and p38 pathways

Figure 3

Praja2 ubiquitylates but does not degrade MFHAS1. (a) HEK293 cells were transiently transfected with plasmids expressing His-tagged MFHAS1, HA-tagged ubiquitin and Flag-tagged praja2 or with an empty PCDH plasmid. After 24 h of transfection, the cells were treated with MG132 (20 μM) for 6 h and were collected. The cell lysates were immunoprecipitated with anti-His antibody. (b) HEK293 cells were transiently transfected with plasmids expressing His-tagged MFHAS1, HA-tagged ubiquitin and Flag-tagged praja2 or Flag-tagged praja2rm or with and an empty PCDH plasmid. After 24 h of transfection, the cells were treated with MG132 (20 μM) for 6 h and were collected. The cell lysates were immunoprecipitated with anti-MFHAS1 antibody. (c) HEK293-MFHAS1 cells were transiently transfected with an empty PCDH plasmid and a plasmid expressing Flag-praja2 or Flag-praja2rm. After 24 h of transfection, the cells were treated with or without MG132 (20 μM) for 6 h and were collected. The cell lysates were immunoblotted with the indicated antibodies

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