Emem growth medium

EMEM growth medium, fetal bovine
serum, penicillin–streptomycin,
and supplements (glutamine and pyruvate) were purchased from Biological
Industries (Beit-Haémek, Israel). Apo-transferrin and PBS pH
7.2 were from Sigma Aldrich. The materials used for synthesis and
work-up procedures were purchased from Sigma Aldrich, Merck, Fluka,
and Frutarom and used upon arrival unless otherwise stated. Deuterated
solvents (Sigma Aldrich isotopes products) with a 99.5% minimum deuteration
were used upon arrival. Silica gel for column chromatography (Silica
Gel 60, 63–200 μm mesh) was obtained from E. Merck Ltd.
Pyrrole was run through a short basic alumina column, and aldehydes
were purified by vacuum distillation before use. HPLC analysis was
performed on a combination of a JASCO organizer, a diode array detector
MD-4010, an autosampler AS-4050, and an RHPLC pump PU-4180. The silica
gel (230–400 mesh) used for column chromatography was obtained
from E. Merck Ltd. A purity of >95% for 3-Ga was determined
with HPLC and UV detection at their Soret band region; a summary of
HPLC results and the HPLC conditions is shown in the Supporting Information.

EMEM growth medium, fetal bovine serum, penicillin-streptomycin, and supplements were purchased from Biological Industries (Beit-Haémek, Israel). MitoTracker Green (MTG), LysoTracker green (LTG), and ER-tracker green (ETG) were obtained from Life Technologies, Rhenium (Jerusalem, Israel). Z-VAD-FMK (#S7023) and necrostatin-1 (#S8037) were purchased from Selleck Chemicals (Houston, TX). The materials used for synthesis and work-up procedures were purchased from Sigma Aldrich, Merck, Fluka, and Frutarom and used as received unless otherwise stated. Deuterated solvents (Sigma Aldrich isotopes products) with a 99.5% minimum deuteration were used as received. Silica gel for column chromatography (Silica Gel 60, 63–200 µm mesh) was obtained from E. Merck Ltd. Pyrrole was run through a short basic alumina column and aldehydes were purified by vacuum distillation before use. Anti-caspase-3 polyclonal (CST#9662), polyclonal anti-PARP (CST#9542), and anti-β-actin monoclonal (CST#4970) antibodies were purchased from Cell Signaling Technology (Danvers, MA). Anti-Cleaved Caspase-3 monoclonal antibody (#ab32042) and anti-rabbit IgG H&L HRP-conjugated (#ab6721) antibodies were from Abcam (Cambridge, MA).