We tested the inhibitory effect of the following compounds at the doses indicated except as otherwise stated, Tetrandrine (TETR 10 μM), Verapamil (VER 100 μM), Curcumin (CUR 75 μM), Bafilomycin (BAF 200 nM), Apilimod (APL 25 μM), Tamoxifen citrate (TMX 10 μM), Raloxifene hydrochloride (RLX 10 μM), Fulvestrant (ICI 182,780; FUL 100 μM) and β-Estradiol (EST 100 μM) were purchased from Sigma-Aldrich. Bapta-AM (BAP 10 μM) and YM201636 (YM 1 μM) and also controls Hydroxycloroquine (HCL 10 μM) endosomal acidification inhibitor and Teicoplanin (TEI 10 μM), an inhibitor of cathepsin L (Zhou et al., 2016 (link)) were purchased from Abcam. Stock solutions were dissolved in DMSO and working solutions were freshly prepared in DMEM 2% fetal bovine serum (FBS) at indicated concentrations. First, we pursued cell viability and cytotoxicity tests of all reagents using the CellTiter 96 Non-radioactive Cell Proliferation Assay (Promega) following the Manufacturer's instructions (Supplementary Fig. S2 , Supplementary Fig. S3 , Supplementary Fig. S4 ). We also studied the cytotoxic activity of the organic solvent DMSO. Based on these experiments we selected optimal non-toxic working concentrations for infection assays.
Ym201636
Manufactured by Abcam
YM201636 is a pharmaceutical compound that acts as a selective inhibitor of the tyrosine kinase enzyme Janus kinase 2 (JAK2). It is commonly used in scientific research to study the role of JAK2 in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Tetrandrine, by Merck Group (1 mentions)
Tamoxifen citrate, by Merck Group (1 mentions)
Apilimod, by Merck Group (1 mentions)
Raloxifene hydrochloride, by Merck Group (1 mentions)
Bafilomycin, by Merck Group (1 mentions)
β estradiol, by Merck Group (1 mentions)
Celltiter 96 non radioactive cell proliferation assay, by Promega (1 mentions)
Curcumin, by Merck Group (1 mentions)
Verapamil, by Merck Group (1 mentions)
Fulvestrant, by Merck Group (1 mentions)
Bapta am, by Abcam (1 mentions)
Lysotracker dnd 99 l 7528, by Thermo Fisher Scientific (1 mentions)
Apilimod, by US Biological (1 mentions)
3 protocols using ym201636
1
Inhibitory Effect of Compounds on Infection
2
Lysosomal Marker Antibody Staining
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Mouse monoclonal anti-LampI (clone H4A3) antibody (sc-2001) was obtained from Santa Cruz Biotechnology. Mouse monoclonal anti-maltose-binding protein (MBP) antibody (E8032) and anti-mouse Alexa 555 secondary antibody (4409S) were purchased from New England Biolabs. Lysotracker DND-99 (L-7528) was purchased from Thermo Fisher. YM201636 was purchased from Abcam (ab141370), Apilimod from US Biological (002800).
3
Optimization of PIKfyve Inhibitor Treatment
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YM201636 was purchased from Abcam (ab141370). Apilimod was purchased from USBiolgical (002800). Both YM201636 and Apilimod were applied at the concentrations and for the times indicated in the figure legends. It was noted that the YM201636 PIKfyve inhibitor easily precipitated in the presence of various transfection reagents. Therefore, cells were washed at least three times with complete medium before YM201636 was applied.
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