Anti egfp

For the preparation of whole-cell extracts, cells were washed with PBS and then lysed in NETN buffer (50 mM Tris-HCl (pH 7.9), 150 mM NaCl, 0.5 mM EDTA, and 0.5% NP40) containing protease and phosphatase inhibitors as described above. Proteins of cell extracts were denatured in sample buffer and further analyzed by SDS-PAGE as described before [45 (link)]. Subsequently, Western blot analysis was performed using anti-HMGCS1 (GeneTex, Irvine, CA, USA and Santa Cruz, CA, USA), anti-Oct4 (GeneTex), anti-SOX-2 (GeneTex), anti-EGFP (GeneTex), anti-eIF2α (GeneTex), anti-p-eIF2α (Cell Signaling), anti-PERK (GeneTex), anti-p-PERK (GeneTex), anti-ATF4 (GeneTex), anti-GCN2 (GeneTex), anti-B23 (Santa Cruz), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH, GeneTex) antibodies.

The following primary antibodies were used in the course of this study: anti-Parvalbumine (EMD Millipore, Billerica, MA, MAB1572, 1:500), anti-eGFP (Genetex, GXT13970, 1:500), anti-PKC (Santa Cruz Biotechnology, Santa Cruz, CA, sc-209, 1:500), anti-humanNT3 (ThermoFisher Scientific, PA1-18385, 1:500), anti-alpha-tubulin (Genetex, Irvine, CA, GTX11304, 1:5000). The following secondary antibodies were used in the course of this study: anti-mouse-Alexa635 (ThermoFisher Scientific, A31574, 1:250), anti-rabbit-Alexa546 (ThermoFisher Scientific, A11081, 1:250), anti-chicken-Alexa488 (ThermoFisher Scientific, A11039, 1:250), anti-Rabbit IgG, HRP conjugated (Promega, W4011, 1:2000), anti-mouse IgG, HRP conjugated (Promega, Madison, WI, W4021, 1:2000).