Supplementary Figure 3: Identification and characterization of p-LLGL2 (Tyr499) and MST1 (Lys59me2).
From: A ROR1–HER3–lncRNA signalling axis modulates the Hippo–YAP pathway to regulate bone metastasis
(a) Summary of top LLGL2-associated proteins identified by LLGL2 pull-down followed by MS in MDA-MB-231 cells. (b and e) Annotated MS/MS spectrum assigned to the LLGL2 peptide VGSFDP[p]YSDDPR, at 717.782 Da. (b) and the STK4/MST1 peptide ETGQIVAI[dime] KQVPVESDLQEIIK, at 1233.200 Da. (e). Data acquired from analysis of the tryptic digest by high-sensitivity LC-MS/MS on an Orbitrap Elite high-resolution mass spectrometer. (c and f) IB detection of p-LLGL2 (Tyr499) (c) and MST1 (Lys59me2) (f) in MDA-MB-231 cells treated with or without NRG1, using antibodies pre-incubated with indicated blocking peptides. Antibodies generated from two independent rabbits were tested. (d) IB detection of indicated proteins in cells transfected with indicated expression vectors followed by NRG1 stimulation. Scanning densitometric analysis was performed for p-LLGL2 (Tyr499) blot. (g) IB detection of indicated proteins in MDA-MB-231 cells treated with LPA or NRG1. Unprocessed original scans of all blots with size marker are shown in Supplementary Fig. 9.
