anti p mlc2 ser 19  (Cell Signaling Technology Inc)

Journal: Cell reports

Article Title: The C. elegans BRCA2-ALP/Enigma Complex Regulates Axon Regeneration via a Rho GTPase-ROCK-MLC Phosphorylation Pathway.

doi: 10.1016/j.celrep.2018.07.049

Figure Lengend Snippet: Figure 6. LET-502, BRC-2, and ALP-1 Promote Axon Regeneration through Phosphorylation of the MLC (A) Structure of MLC-4. Schematic diagrams of MLC-4 and its mammalian counterpart MLC2 are shown. Asterisks indicate regulatory residues Thr-17 and Ser-18. EF-hand domains (EFh; black) are shown. (B) Percentages of axons that initiated regeneration 24 hr after laser surgery. Error bars indicate 95% CIs. *p < 0.05, as determined by Fisher’s exact test. NS, not significant. (C) Yeast two-hybrid assays for the interaction of ALP-1 with MLC-4. The re- porter strain PJ69-4A was co-transformed with expression vectors encoding GAL4 DBD-ALP-1 and GAL4 AD-MLC-4 (wild-type [WT], DD, or AA), as indi- cated. Yeasts carrying the indicated plasmids were grown on a selective plate lacking histidine and containing 1 mM 5-AT for 4 days.

Article Snippet: The permeabilized worms were incubated with buffer A (20 mM Tris-HCl [pH 8.0], 150 mMNaCl, 1% Triton X-100, 1 mM EDTA, 3%BSA, and 0.05% sodium azide) overnight at 4 C. The worms were then incubated with primary antibodies (3200dilutedwithbufferA) overnight at 4 C.Theprimary antibodieswere anti-phospho-MLC2 Ser-19 rabbit antibody (#3671, CST) and anti-GFP mouse monoclonal antibody 1E4 (M048-3, MBL International).

Techniques: Phospho-proteomics, Transformation Assay, Expressing