TrueBlot^(R) Anti Ig  

Western Blot of TrueBlotR Anti-Rabbit Ig IP Agarose Beads. Lane 1: Protein Standard Opal Pre-stained (p/n MB-210-0500). Lane 2: Rb-a-GFP Input. Lane 3: Rb-a-GFP Unbound. Lane 4: Rb-a-Elute. Primary Antibody: TrueBlotR Anti-Rabbit Ig IP Agarose Beads (00-8800-25) at 1 μg/mL for overnight at 4？C. Secondary Antibody: TrueBlotR Rabbit Secondary HRP Antibody (p/n 18-8816-33) at 1:10,000 for 30 min at RT. Block: MB-070 at RT for 30 min. Predicted/Observed size: 27 kDa.

ADGRF1 overexpression activates Gαs/Gαq pathways. BT474 clone 1 and SKBR3 clone 2 were grown in absence (−) or presence (+) of doxycycline (Dox). A, Co-immunoprecipitation with anti-Gαs and immunoblotting with anti-HA indicate that ADGRF1 couples to Gαs in both BT474 and SKBR3. B, Basal levels of cAMP were significantly increased upon ADGRF1 overexpression with Dox in both cells. C, Co？immunoprecipitation with anti-Gαq and immunoblotting with anti-HA indicate that ADGRF1 couples also to Gαq in both cells. D, Basal levels of IP1 were significantly increased upon ADGRF1 overexpression with Dox in BT474 but not SKBR3 cells. *indicates statistically significant difference？P？< .05 by unpaired？t？test (N = 3？4). FIGURE 5. PMID: 34110646.

MLL5 interacts with CRX via its CD4 domain(A) Interaction between endogenous MLL5 and CRX. Equivalent amounts of retinal lysates from？Mll5-WT or？Mll5-KO mice were immunoprecipitated with anti-MLL5 antibody, anti-CRX or normal rabbit IgG, followed by western blotting detection.(B) Schematic representation of MLL5 truncated mutants. PS, PHD/SET domain; CD, central domain; CT, C-terminal domain. ΔCT4, deleted CT4 domain.(C) MLL5 central domain (MLL5-CD) binding to CRX. HEK293T cells expressing HA-CRX together with Flag-MLL5-PS, Flag-MLL5-CD, or Flag-MLL5-CT were subjected to immunoprecipitation with the indicated antibodies followed by western blotting detection.(D) Interaction between MLL5-CD4 and CRX. HEK293T cells expressing HA-CRX and Flag-MLL5-CD4 were immunoprecipitated with anti-HA antibodies and detected by anti-Flag and anti-HA antibodies.(E) Depletion of MLL5 CD4 compromises MLL5 interaction with CRX. HEK293T cells expressing HA-CRX along with Flag-MLL5-FL or Flag-MLL5-ΔCD4 were immunoprecipitated with anti-HA and anti-Flag antibodies followed by western blotting detection.(F) MLL5-ΔCD4 was less potent than MLL5-WT at rescuing transcription of？Rho.？qPCR analysis of？Rho？transcript in 661W (CRX OE) cells transfected with MLL5-siRNA (siMLL5) followed by transduction with the indicated lentivirus. Rho expression was normalized to the expression of Tbp. Error bars represent SEM (n？= 3), ？？p？< 0.01, Student’s t test. Results in (A？F) are representative of at least three experimental repeats. Fig 4. PMID: 35359806.