Rabbit anti carm1

The primary antibodies used and the dilutions were as follows: guinea pig anti-insulin (1:50; Dako), mouse anti-glucagon (1:200; Abcam), mouse anti-actin (1:800; Fitzgerald), mouse anti-Smn (1:5000; BD Transduction Laboratories), rabbit anti-phospho-AKT (Ser473) (1:500; Cell Signaling), rabbit anti-AKT (1:500; Cell Signaling), rabbit anti-phospho-CREB (Ser133) (1:500; Cell Signaling), rabbit anti-CREB (1:500; Cell Signaling), rabbit anti-Pck1 (1:1000; Abcam), rabbit anti-CARM1 (1:5000; Bethyl Laboratories) mouse anti-2H3 (neurofilament 165 kDa, 1:100; Hybridoma Bank) and mouse anti-SV2 (1:250; Hybridoma Bank). The secondary antibodies used were as follows: donkey anti-guinea pig biotin-SP-conjugated (1:200; Jackson Immuno Research), streptavidin-Cy3-conjugated (1:600; Jackson Immuno Research), Alexa Fluor 488 goat anti-mouse (1:500; Molecular Probes), HRP-conjugated goat anti-rabbit IgG (1:5000; Bio-Rad) and HRP-conjugated goat anti-mouse IgG (1:5000; Bio-Rad).

Simultaneous RNA ISH and immunofluorescence were performed as described (Yin et al. 2012 (link)) with slight modifications. Hybridization was performed with in vitro transcribed Dig-labeled probes. For colocalization studies, cells were costained with mouse anti-p54^nrb (BD) and/or rabbit anti-CARM1 (Bethyl Laboratories). The nuclei were counterstained with DAPI. Images were taken with a Zeiss LSM 510 microscope or an Olympus IX70 DeltaVision RT deconvolution system microscope. For statistical analysis, >300 cells from each group were observed and calculated. Image analyses of signal intensity were carried out by Image-Pro Plus according to standard protocols and were described previously (Yin et al. 2015 (link)).