Skip to main content
. Author manuscript; available in PMC: 2021 Aug 20.
Published in final edited form as: J Cell Physiol. 2018 Dec 19;234(8):13220–13232. doi: 10.1002/jcp.27994

FIGURE 4.

FIGURE 4

BRAF stabilizes ERK3 protein without requiring BRAF kinase activity. (a) Western blot analysis analysis of ERK3 level by BRAF (WT), BRAFV600E and BRAF kinase-dead (KD) plasmids transiently overexpression for 2 days in (a) nonmelanoma cell 293T or (b) A375. All three BRAF constructs are all able to induce ERK3 levels compared with pcDNA3 control in both cell lines. Immunoblots are quantified to the right. (c) BRAF-KD overexpression in 293T results in ERK3 protein stabilization. CHX treatment was performed at 2 days posttransfection of pcDNA3 control or BRAF-KD. Western blots and degradation trend line are representatives of multiple experiments expressed as a mean ± standard deviation (n = 3). β-Actin or GAPDH serves as a loading control. CHX: cycloheximide; ERK3: extracellular signal-regulated kinase 3; GAPDH: glyceraldehyde 3-phosphate dehydrogenase