Human NRG1 Isoform SMDF Antibody

Detection of Neuregulin-1 Isoform SMDF by Western Blot

SMDF transforms via ErbB receptor signalling.(A) Western blot analysis of indicated proteins in NSLT cells expressing SMDF, SMDF* (point mutant incapable of binding ErbB receptors) or control vector (LXSN), and SMDF(T) cells derived from SMDF tumours in the absence or presence of the ErbB-2 inhibitor, 4557W (40 nM). (B) Cell counts of triplicate wells in the presence or absence of the indicated concentrations of 4557W −/+ S.D.. Figure is representative of 3 independent experiments. (C) Number of colonies formed in soft agar assays of NSLT cells expressing SMDF, Ras or control vector LXSN in the absence or presence of the indicated inhibitors. U0126 (UO), LY294002 (LY) and the ErbB inhibitor (ErbBI). 12 fields were counted per well with the results shown as average/well −/+ S.D.. Results are representative of two separate experiments (D) Soft agar assays of NSLT cells infected with SMDF, SMDF* or control vector (E) 105 NSLT cells expressing SMDF, Ras or control vector (LXSN) were seeded in triplicate into 6 well dishes and counted at 72 h. The inhibitors U0126 (20 µM), LY294002 (20 µM) or control vehicle were added where indicated at 36 hours. Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN). (F) Western blot analysis of total lysates from NSLT cells expressing SMDF or control vector (LXSN) collected 3 (preconfluent), 6 and 9 days after seeding. Plates were medium changed daily. (G) Western blot analysis of total lysates from NSLT cells expressing SMDF, Ras or control vector (LXSN) in attached (att) or anchorage-independent conditions (sus). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20668675), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Neuregulin-1 Isoform SMDF by Western Blot

SMDF promotes oncogenic transformation.(A) Western blot analysis of total lysates from NSLT cells selected to express human SMDF or control vector (LXSN). SMDF(T) are derived from tumours, induced by injecting NSLTSMDF cells into nude mice. (B) Cell counts −/+ S.D. following seeding of 105 NSLT cells per well expressing SMDF, Ras or control vector (LXSN) in triplicate in 6 well plates. Medium was changed daily. Experiment shown is representative of 3 separate experiments. Representative phase-contrast images of the cells at high density. (C) NSLT cells expressing SMDF, Ras or control vector (LXSN) were injected into the flanks of nude mice. Ras cells were only injected into a single flank. Picture shows examples of tumours formed (SMDF (left), Ras (right)) and the graph shows the average growth of the tumours −/+ S.D.. (D) Representative images of cells dissociated from NSLTSMDF derived tumours stained for LT (upper panel) or the Schwann cell marker, S100 (lower panel). Primary Schwann cells (NS) were infected with retroviral vectors expressing SMDF, Ras or the control vector (LXSN) and selected in G418. One week following selection, the cultures were fixed and stained for the senescence marker beta -Galactosidase at pH 6 (E) or seeded in triplicate and counted for four passages (F). Western blot shows p-ERK levels in lysates prepared from NS cells expressing Ras, SMDF or control vector (LXSN) six days after infection. (G) 2×105 NS cells infected with SMDF or control vector (LXSN) were plated in triplicate in 6 well dishes and counted at the indicated days. Medium was changed daily. Error bars show −/+ S.D.. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20668675), licensed under a CC-BY license. Not internally tested by R&D Systems.