Monoclonal anti rhodopsin1

All fluorescence images were obtained with a Zeiss LSM710 confocal microscope using a 20X objective lens. The following antibodies were used: rabbit anti-cleaved caspase-3 (diluted 1:50; #9661, Cell Signaling, MA, USA), monoclonal anti-rhodopsin1 (diluted 1:500; Developmental Studies Hybridoma Bank, University of Iowa, IA, USA), rabbit anti-dsRed (diluted 1:500; Clontech Laboratories, Inc., CA, USA), and guinea pig anti-ATF4 (diluted 1:500^{32 (link)}). In addition, Apoptosis was analyzed by TUNEL staining using an ApopTag Red In Situ Apoptosis Detection kit (S7165, Merck Millipore, MA, USA).

All fluorescent images were obtained with a Zeiss LSM710 confocal microscope, using a ×20 or x40 objective lens. The following antibodies were used: rabbit anti-dsRed antibody (1:50 from Clontech or 1:500 from Dr. S.W. Kang), rabbit anti-GFP (1:2000, Molecular Probes, catalogue no. A6455), mouse anti-armadillo (1:500; Developmental Studies Hybridoma Bank, N2 7A1, University of Iowa, USA), monoclonal anti-rhodopsin1 (1:500; Developmental Studies Hybridoma Bank, 4C5, University of Iowa, USA), actin-phalloidin (1:200; Molecular Probes). To generate the guinea-pig anti-ATF4 antibody, the full length of the ATF4 coding sequence was subcloned into XhoI and NotI sites in pET14b (Novagen). The resulting ~50 kDa His-tagged recombinant protein was purified to generate a polyclonal antibody. The antisera were subsequently affinity purified against the same epitope [19 (link)].