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. 2021 Jul 9;7:91. doi: 10.1038/s41523-021-00300-1

Fig. 3. JAB1 is an MK2 substrate and is required for robust AP1 activity in TNBC cells.

Fig. 3

a MDA-MB-231 cells were immunoprecipitated with either JAB1 or MK2 antibody and the immunoprecipitates were subjected to western blot to detect MK2 or JAB1 respectively. All blots derived from the same experiment and were processed in parallel. b MDA-MB-231 cells were subjected to immunofluorescence staining to detect JAB1 and MK2. Data are the representative of four independent experiments. c Diagram of HA-tagged full-length JAB1, N-terminal Jab1 (JAB1-N), and C-terminal JAB1 (JAB1-C). d HEK293 cells were transfected with empty plasmid (Control) or with plasmids encoding HA-tagged full-length JAB1 (FL), JAB1-N or JAB1-C, and cell lysates were immunoprecipitated with MK2 pAb. Immunoprecipitates were subjected to western blot analysis to detect interactions between the JAB1 proteins and MK2 by HA mAb. Data are the representative of two independent experiments. All blots derive from the same experiment and were processed in parallel. e BT549, MDA-MB-231 or MDA-MB-436 cells were treated with 30 nM JAB1 siRNA (siJAB1-1 and siJAB1-2) or scrambled siRNA control for 3 days followed by analyzing AP 1 activity. Data are means ± SD from three experiments. *P < 0.01 vs Control. f Wild-type MEFs were first transduced with MKK3D, MKK6D, MK2-CA, or empty vector (Control) for 3 days and then treated with 30 nM JAB1 siRNAs for 3 days followed by analyzing AP1 activity. Data are means ± SD from three experiments. *P < 0.01 vs Control.