TrueBlot^(R) Anti Ig  

Immunoprecipitation/Western Blot using GFP Protein. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500). Lane 2: GFP Input (p/n 000-001-215) Reduced [10μL]. Primary IP Antibody: Mouse Anti-GFP (p/n 600-301-215) at 10μg overnight at 2-8°C. Secondary Antibody: TrueBlot Anti-Mouse Ig IP Agarose Beads (p/n 00-8811-25) at 500μg for 1hr at RT. Buffer: BlockOut Buffer (p/n MB-073) for 30 mins at RT. Exposure: 7 sec.

Loss of LIFR activates NF-κB signaling through SHP1, leading to upregulation of LCN2. (a)？HEK293T cells were transfected with HA-FLAG-SHP1 and SFB-tagged GFP or LIFR. LIFR-SFB protein was pulled down with S-protein beads, followed by immunoblotting with antibodies against FLAG and HA.？(b)？HEK293T cells were transfected with MYC-SHP2 and SFB-tagged GFP or LIFR. LIFR-SFB protein was pulled down with S-protein beads, followed by immunoblotting with antibodies against FLAG and MYC.？(c)？HEK293T SFB-GFP and SFB-LIFR stable cell lines were infected with the scrambled (Scr) or sh-SHP1 lentivirus, followed by transfection with a K63-specific mutant of His-Xpress-ubiquitin (Ub). 48？h later, cells were subjected to pulldown with nickel beads and immunoblotting with antibodies against TRAF6 and Xpress.？(d)？Control and LIFR-overexpressing PLC/PRF/5 cells were transduced with SHP1 shRNA and immunoblotted with the indicated antibodies.？(e)？Control (Scr) and LIFR-knockdown HEK293T cells were transfected with FLAG-TRAF6. 48？h later, cells were immunoprecipitated with a FLAG-specific antibody and immunoblotted with antibodies against LIFR, SHP1, and FLAG. Fig. 4. PMID: 34921145.

GAGE12 modulates H3K56Ac through attenuation of HDAC activity.(E) Interaction of FLAG-GAGE12, SYNM, HDAC1/2, and β-actin in total nuclear lysates of mock and f:G12-OE cells. In FLAG immunoprecipitates, SYNM, HDAC1/2, and β-actin are only pulled down in the presence of FLAG-GAGE12 (lanes 3 and 4). In SYMN immunoprecipitates, SYNM only pulled down HDAC1 and HDAC2 in the presence of FLAG-GAGE12. SYNM interacts with β-actin, even in the absence of GAGE12 (lanes 5 versus 6). (F) Increased HDAC1-HDAC2-β-actin interaction in the presence of GAGE12. In HDAC1 and HDAC2 immunoprecipitates, increased β-actin was found in the pull-down lysates only in the presence of GAGE12. Figure？6. PMID: 34469741.