Biliverdin

Cell images were taken with a Leitz-Wetzlar Type 307 microscope equipped with a 12-bit CCD camera and a Nikon Eclipse Ti2 microscope. Fluorescence channels were merged using ImageJ software. Timelapse imaging was conducted with HeLa cells stably expressing GFP-tagged GalT and HeLa cells stably expressing GFP-tubulin on an Olympus FluoView1000 using a 20x/0.95 NA water objective. Z-stack slices were optimized for the objective at 1.18 mm thickness. For the duration of the imaging, cells were on a 37 °C heat plate and maintained in L-15 media (Thermo Scientific, Waltham, MA) supplemented with 10% fetal bovine serum. Cells transfected with iRFP-tagged plasmids were incubated with 5 μM biliverdin (Cayman Chemical, Ann Arbor, MI) for 30 min prior to imaging.

Grounded powder of Arthrospira platensis (lot number D1246) was purchased from Abundance (Kfar Rupin, Israel). Biliverdin (CAS registry number 856699-18-8) purity >98% was purchased from Cayman chemical company (Ann Arbor, MI, USA). Sodium phosphate dibasic, sodium phosphate monobasic, sodium citrate dihydrate, and Tris (hydroxymethyl) aminomethane were purchased from Spectrum Chemical (New Brunswick, Canada). Hydrochloric acid, sodium hydroxide, citric acid, methanol absolute CP, ethanol absolute AR, and dichloromethane CP were purchased from Bio-lab (Jerusalem, Israel). Ammonium persulfate was purchased from Sigma Aldrich (St. Louis, MO, USA). HPLC grade acetonitrile was purchased from J.T Baker (Gilwice, Poland). HPLC grade water was purchased from Marcon (Giliwice, Poland).

The fetal rhesus kidney (FRhK-4) cell is a HAV-permissive cell line derived from the rhesus monkey. Dulbecco’s modified Eagle’s medium (DMEM) containing 4% heat-inactivated fetal bovine serum (FBS, Gibco, Campinas, Brazil) and antibiotic-antimycotic reagent (Gibco, Campinas, Brazil) was used to maintain this cell line under a 5% CO₂ atmosphere at 37 °C.
The HAV strain HM-175/18f, clone B (VR-1402) was obtained from the American Type Culture Collection. Ten genomic equivalents (GE) copies of the HAV per cell were used to inoculate FRhK-4. Cells inoculated with the virus were maintained in DMEM supplemented with 4% heat-inactivated FBS and antibiotic–antimycotic reagent.
We purchased hemin, cobalt protoporphyrin IX (CoPP-9), andrographolide, zinc protoporphyrin IX (ZnPP-9), iron (III) chloride (FeCl₃), and carbon monoxide-releasing molecule (CORM-3) from Sigma-Aldrich (St. Louis, MO, USA). Biliverdin was purchased from Cayman Chemical (Ann Arbor, MI, USA).