All chemicals were purchased from Sigma-Aldrich unless otherwise noted. Py–Im polyamides were synthesized by microwave-assisted, solid-phase synthesis on Kaiser oxime resin (Novabiochem) according to previously described protocols (Baird & Dervan, 1996 ; Puckett et al. 2012 (link)). Polyamide 5 was synthesized on hydrazine resin (855037, Novabiochem) in the same manner as the other polyamides and cleaved from resin in the same manner after 10 min oxidation of the hydrazine by Cu(II)SO4 in pyridine and DMF. Polyamides were purified by reverse-phase HPLC and lyophilized. EZ-Link NHS-PEG4-biotin (Pierce) was conjugated in 5% Hünig’s base in DMF. Purity and identity of compounds were verified by analytical HPLC and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (SI Table 2 ).
Brefeldin A was purchased from BD Biosciences, 7-AAD from eBiosciences and Z-VAD-fmk from Promega. Antibodies purchased from Abcam are: polyclonal rabbit anti-calreticulin (ab2907), polyclonal rabbit anti-ERp57 (ab10287), polyclonal goat anti-rabbit Alexa Fluor 488 (ab150081). Antibodies from Life Technologies are: anti-Annexin V Alexa Fluor 488 conjugate (A13201) and mouse monoclonal anti-actin (AM4302). Antibodies from Cell Signaling Technologies are: rabbit polyclonal anti-PARP (9542) and rabbit monoclonal anti-phospho-H2AX (ser139, 9718).
Brefeldin A was purchased from BD Biosciences, 7-AAD from eBiosciences and Z-VAD-fmk from Promega. Antibodies purchased from Abcam are: polyclonal rabbit anti-calreticulin (ab2907), polyclonal rabbit anti-ERp57 (ab10287), polyclonal goat anti-rabbit Alexa Fluor 488 (ab150081). Antibodies from Life Technologies are: anti-Annexin V Alexa Fluor 488 conjugate (A13201) and mouse monoclonal anti-actin (AM4302). Antibodies from Cell Signaling Technologies are: rabbit polyclonal anti-PARP (9542) and rabbit monoclonal anti-phospho-H2AX (ser139, 9718).