Uranyl acetate

Small pieces (~1 mm³) of the left ventricle were fixed at 4 °C in 4% buffered glutaraldehyde (SERVA, Frankfurt, Germany) with 0.2 M cacodylate buffers (TAAB essential for microscopy, Aldermaston, Berks, UK) and processed for TEM as we previously described [22 (link)]. Contrasted ultrathin sections with uranyl acetate and lead citrate (Loba Chemie Pvt. Ltd., Colaba, Mumbai, India) were examined and photographed using a Philips EM 208S TEM (FEI Company, Eindhoven, The Netherlands) at an accelerating voltage of 80 kV.

Ethanol, mEthanol, chloroform, glacial acetic acid, HNO₃, H₂O₂, Na₂HPO₄, NaH₂PO₄, AgNO₃, Na₂CO₃ and sodium dodecyl sulphate (SDS) were purchased from Merck, Bengaluru, India. Bovine serum albumin and chitosan powder were purchased from Sisco Research Laboratories, Chandigarh, India. Glutaraldehyde was purchased from TCI Chemicals, Chandigarh, India. A Spurr resin kit and OsO₄ were purchased from Electron Microscopy Sciences, Delhi, India. Uranyl acetate was purchased from LobaChemie, India. CuCl, selenourea, oleylamine, citric acid, sodium citrate monobasic, lead citrate, Laemmli buffer, ammonium persulfate, trizma base, tetramethylethylenediamine (TEMED), acrylamide, N,N′- methylenebis(acrylamide), 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), KOH, phenylmethylsulfonyl fluoride (PMSF) cocktail, KH₂PO₄, K₂HPO₄, glycerol, and formaldehyde were purchased from Sigma-Aldrich, Bengaluru, India. Bradford reagent, macerozyme R-10, 1X phosphate buffer saline (PBS), sodium thiosulfate, and ascorbate were purchased from HiMedia, Mumbai, India. Rehydration buffer, IPG strips of pH range 3–10, a ReadyPrep 2D starter kit equilibration buffer I and II, and a ReadyPrep 2D clean-up kit was purchased from Bio-Rad, Gurugram, India. A precision plus protein kaleidoscope prestained protein ladder was purchased from Bio-Rad.

All the routinely used biochemicals were of molecular biology grade and procured from MERCK or similar sources unless otherwise stated. Cell culture reagents were from Lonza and Thermo Fisher Scientific. Restriction enzymes and DNA modifying enzymes were from New England Biolabs and Thermo Fisher Scientific. DNA isolation kits were from QIAGEN and MACHEREY-NAGEL. ProLong Diamond Antifade Mountant, Hoechst, DAPI, SuperSignal Chemiluminescent substrate and 2’,7’-dichlorodihydrofluorescein diacetate acetyl ester (H₂DCFDA) were from Thermo Fisher Scientific. Secondary antibodies were from Thermo Fisher Scientific and Jackson ImmunoResearch. Primary antibodies were from Cell Signalling Technologies, Thermo Fisher Scientific, MERCK and ChromoTek. WR99210 was a kind gift from David Jacobus (Jacobus Pharmaceutical, Princeton, U.S.A.). Protease inhibitor cocktail, trimethoprim and N-acetylglucosamine (GlcNAc) were from MERCK. Glutaraldehyde, formaldehyde, osmium tetroxide, lead citrate solution, and sodium cacodylate buffer were from Electron Microscopy Sciences. Uranyl acetate was from Loba Chemie and epoxy resin constituents were from TED PELLA. P. falciparum 3D7 and D10 strains were obtained from the Malaria Research and Reagent Reference Resource centre (MR4). All the plastic ware was from standard manufacturers such as Corning Inc, Nalgene, Thermo Fisher Scientific and Tarsons.