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Apilimod

Manufactured by MedChemExpress
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Apilimod is a laboratory product manufactured by MedChemExpress. It is a small molecule compound that can be used for research purposes.

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Lab products found in correlation

Goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Ca 074, by MedChemExpress (1 mentions) Bafilomycin a, by Merck Group (1 mentions) Mouse monoclonal anti flag m2 antibody, by Merck Group (1 mentions) Nh4cl, by Merck Group (1 mentions) Ym201636, by MedChemExpress (1 mentions) Anti ifn γ xmg1, by BioXCell (1 mentions) Anti nk1.1 pk136, by BioXCell (1 mentions) Anti cd8 2.43, by BioXCell (1 mentions) Anti cd19 1d3, by BioXCell (1 mentions) Anti cd4 yts 177, by BioXCell (1 mentions) Il 12, by STEMCELL (1 mentions) Ifn γ, by STEMCELL (1 mentions) Okadaic acid, by Santa Cruz Biotechnology (1 mentions) Mouse anti gapdh, by Thermo Fisher Scientific (1 mentions) Torin 1, by Cayman Chemical (1 mentions) Rabbit anti akt, by Cell Signaling Technology (1 mentions) Rabbit anti ulk1, by Cell Signaling Technology (1 mentions) Fk506, by Cayman Chemical (1 mentions) Rabbit anti 4e bp1, by Cell Signaling Technology (1 mentions)

11 protocols using apilimod

1

Biochemical Reagents for COVID-19 Research

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Broad-spectrum cysteine protease inhibitor E64D (#HY-100229), Cathepsin L-specific inhibitor SID 26681509 (#HY-103353), Cathepsin B-specific inhibitor CA-074 (#HY-103350), PIKfyve inhibitors apilimod (#HY-14644) and YM201636 (#HY-13228), were purchased from Med Chem Express (MCE, New Jersey, USA). Calcium channel blocker tetrandrine (#S2403) was purchased from Selleck Chemicals (Texas, USA). Endosome acidification inhibitors NH4Cl (#A9434) and bafilomycin A (#19-148) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit polyclonal against SARS S1 antibodies (#40150-T62), mouse monoclonal against MERS-CoV S2 antibody (#40070-MM11), mouse monoclonal against SARS S1 antibody (#40150-MM02), rabbit polyclonal against HIV-1 Gag-p24 antibody (11695-RP01) were purchased from Sino Biological Inc. (Beijing, China). Goat polyclonal against human ACE2 antibody (#AF933) was purchased from R&D Systems (Minnesota, USA). Mouse monoclonal anti-FLAG M2 antibody was purchased from Sigma-Aldrich. Donkey Anti-Rabbit IgG (#711-035-152), Goat Anti-Mouse IgG (#115-035-146), and Rabbit Anti-Goat IgG (#305-035-003) were purchased from Jackson ImmunoResearch (West Grove, PA, USA). Alexa Fluor 488-conjugated goat anti-rabbit IgG, Alexa Fluor 488-conjugated goat anti-mouse IgG were purchased from ZSGB-Bio LLC (Beijing, China).
Ou X., Liu Y., Lei X., Li P., Mi D., Ren L., Guo L., Guo R., Chen T., Hu J., Xiang Z., Mu Z., Chen X., Chen J., Hu K., Jin Q., Wang J, & Qian Z. (2020). Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV. Nature Communications, 11, 1620.
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2

Modulating Immune Responses in Viral Infection

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Agonistic mouse anti-CD40 (clone FGK4.5/FGK45, BioXCell, Lebanon, NH, USA), anti-CD154/CD40L (MR-1, BioXCell), anti-NK1.1 (PK136, BioXCell), anti-CD19 (1D3, BioXCell), anti-IL12 (R1-5D9, BioXCell), and anti-IFNγ (XMG1.2, BioXCell) antibodies were diluted to 200 µg in PBS and administered 24 h prior to viral challenge. Anti-CD4 (YTS 177, BioXCell) and anti-CD8 (2.43, BioXCell) antibodies were diluted to 200 µg/100 µL in PBS, pooled to a total of 400 µg (200 µL), and administered 24 h prior to downstream use. Isotype IgG controls (BioXCell) were used at equivalent doses in all experiments. All antibodies were administered in a final volume of 100–200 µL via intraperitoneal (i.p.) injection. Cytokines used in this study include IL-12 (StemCell Technologies, Vancouver, CA, USA, catalog #78028.1) and IFN-γ (StemCell Technologies, catalog #78020.1), which were given at 5 µg in 100 µL of PBS via i.p. injection 24 h prior to viral challenge. For the disruption of IL-12 production, the inhibitor apilimod (MedChemExpress, Monmouth Junction, NJ, USA, catalog #HY-14644) was administered via i.p. injection at a concentration of 2.5 mg/kg. The drug was administered on day −1, day 0, and day 1 post-infection with rVSV-EBOV GP.
Rogers K.J., Richards P.T., Zacharias Z.R., Stunz L.L., Vijay R., Butler N.S., Legge K.L., Bishop G.A, & Maury W. (2023). CD40 Signaling in Mice Elicits a Broad Antiviral Response Early during Acute Infection with RNA Viruses. Viruses, 15(6), 1353.
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3

Apilimod Modulates Lysosomal Dynamics

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Apilimod (HY-14644) was purchased from MedChem Express (Monmouth Junction, NJ). Aliquots were prepared in ultra-pure grade dimethyl sulfoxide (DMSO) (VWR; 97063-136) and stored at −20°C until use. On each experimental day, a fresh aliquot of Apilimod (200 μM) was thawed. At 24 hours after transfection, cells were treated with either Apilimod (final concentration, 200 nM) or DMSO (1:2000, v/v) for 1 to 1.5 hours at 37°C. Cells were prepared for live cell imaging. As a positive control for Apilimod treatment, the diameter of lysosomal-associated membrane protein 1 (LAMP-1)–positive intracellular vesicles was measured and found to be substantially increased in treated compared to untreated cells (2.2. μm versus 1.7 μm), which is consistent with previous reports (fig. S2L) (31 (link)).
Norris M.J., Husby M.L., Kiosses W.B., Yin J., Saxena R., Rennick L.J., Heiner A., Harkins S.S., Pokhrel R., Schendel S.L., Hastie K.M., Landeras-Bueno S., Salie Z.L., Lee B., Chapagain P.P., Maisner A., Duprex W.P., Stahelin R.V, & Saphire E.O. (2022). Measles and Nipah virus assembly: Specific lipid binding drives matrix polymerization. Science Advances, 8(29), eabn1440.
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4

Antibody Validation and Signaling Pathway Analysis

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The following antibodies were used: rabbit anti-p70S6K (Cell Signaling Technologies; #2708), rabbit anti-p-p70S6K (Thr389) (Cell Signaling Technologies; #9205), rabbit anti-Akt (Cell Signaling Technologies; #9272), rabbit anti-p-Akt (Thr308) (Cell Signaling Technologies; #13038), rabbit anti-p-Akt (Ser473) (Cell Signaling Technologies; #4051), rabbit anti-ULK1 (Cell Signaling Technologies; #8054), rabbit anti-p-ULK1 (Ser757) (Cell Signaling Technologies; #6888), rabbit anti-TFEB (Cell Signaling Technologies; #4240), rabbit anti-p-TFEB (Ser211) (Cell Signaling Technologies; #37681), rabbit anti-4E-BP1 (Cell Signaling Technologies; #9644), rabbit anti-p-4E-BP1 (Thr37/46) (Cell Signaling Technologies; #2855), rabbit anti-mTOR (Cell Signaling Technologies; #2983), rabbit anti-Raptor (Cell Signaling Technologies; #2280), rabbit anti-HA (Cell Signaling Technologies; #3724), mouse anti-α-tubulin (Thermo; 236-10501), mouse anti-GAPDH (Thermo; AM4300), mouse anti-DYKDDDDK (WAKO; clone 1E6), mouse anti-LAMP1 (Development Studies Hybridoma Bank; H4A3), mouse anti-PIKfyve (Development Studies Hybridoma Bank; 3C9), rabbit anti-TFE3 (Sigma; HPA023881), mouse anti-PP2A-Cα/β (Santa Cruz; sc-80665), and rabbit anti-PPP2CA (Proteintech; 13482-1-AP). Apilimod was from MedChem Express. Okadaic acid was from Santa Cruz. Torin1, YM201636, VPS34-IN1, and FK-506 were from Cayman Chemical.
Hasegawa J., Tokuda E., Yao Y., Sasaki T., Inoki K, & Weisman L.S. (2022). PP2A-dependent TFEB activation is blocked by PIKfyve-induced mTORC1 activity. Molecular Biology of the Cell, 33(3), ar26.
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5

Vacuolin-1 and Apilimod Inhibit Lysosome Function

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Vacuolin-1 (18 (link)) was custom synthesized; Apilimod (HY-14644) was from MedChem Express, IN1 was a kind gift from N. Gray, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA (33 (link)), U-18666A (10009085) and Filipin III (70440) were from Cayman Chemical, Bafilomycin A1 (B1793-2UG) was from Sigma-Aldrich, Cycloheximide (239764) was from Calbiochem, and wheat germ agglutinin conjugated with Alexa Fluor-647 (W32466) was from ThermoFisher.
Kang Y.L., Chou Y.Y., Rothlauf P.W., Liu Z., Soh T.K., Cureton D., Case J.B., Chen R.E., Diamond M.S., Whelan S.P, & Kirchhausen T. (2020). Inhibition of PIKfyve kinase prevents infection by Zaire ebolavirus and SARS-CoV-2. Proceedings of the National Academy of Sciences of the United States of America, 117(34), 20803-20813.
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6

Apilimod Inhibits Calvarial Defect Repair

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In brief, 1.0 mm calvarial defects were created in the parietal bones of mice (8 weeks old, n = 4-5 per group). Apilimod (7 mg/kg body weight; MedChem Express, Princeton, NJ, USA) was orally administered twice a week, starting 3 days before surgery, to inhibit the production of IL-12 and IL-23. The mice were euthanized at 4 weeks after the surgery. The parietal bones were harvested and fixed for 24 h in 4% PFA for further study. Investigators were blinded to the treatment group of each sample.
Xu J., Li J., Hu Y., Dai K., Gan Y., Zhao J., Huang M, & Zhang X. (2020). IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders. Theranostics, 10(9), 3925-3938.
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7

Bone Regeneration with Stem Cells and Cytokines

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C57BL/6J, nude mice, IL-12p35-/- mice, or IL-12p40-/- mice (n=4-5 per group, male, 8-10 weeks old) were randomly divided into different groups as detailed in Results. A total of 3.0 × 106 BMMSCs was mixed with 45 mg of β-tricalcium phosphate (β-TCP) ceramic particles (Shanghai Bio-lu Biomaterials Co., Ltd., Shanghai, China) and subcutaneously implanted into the dorsal surface of mice. For in vivo evaluation of the effects of sustained IL-12 and IL-23 release, HyStem-HP Hydrogel (250 μL; ESI BIO stem cell solutions, Alameda, CA, USA) was covered on the surface of the implants containing 200 ng of IL-12 and 200 ng of IL-23 (Peprotech, Rocky Hill, New Jersey, USA). Apilimod (7 mg/kg body weight 20 (link); MedChem Express, Monmouth Junction, NJ, USA) and vehicle were orally administered twice a week, starting three days before surgery, to determine the effect of IL-12/23 inhibitor on bone formation in vivo. The samples collected at 56 days were fixed in 4% paraformaldehyde (PFA) and decalcified with 12.5% ethylenediaminetetraacetic acid (EDTA; pH 7.0).
Xu J., Li J., Hu Y., Dai K., Gan Y., Zhao J., Huang M, & Zhang X. (2020). IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders. Theranostics, 10(9), 3925-3938.
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8

Vacuolin-1 Synthesis and Apilimod Procurement

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Vacuolin-1 REF TK was custom synthesized; Apilimod (HY-14644) was purchased from MedChem Express.
Sanyal A., Scanavachi G., Somerville E., Saminathan A., Nair A., Oikonomou A., Hatzakis N.S, & Kirchhausen T. (2024). Constitutive Endolysosomal Perforation in Neurons allows Induction of α-Synuclein Aggregation by Internalized Pre-Formed Fibrils. bioRxiv.
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9

Antiviral Compound Sourcing and Purity Verification

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The following chemicals were purchased from the indicated companies: bafilomycin A 1 (Cayman Chemical Company, Ann Arbor, MI, USA; 11038), dynasore (Sigma-Aldrich, St. Louis, MO, USA; D7639), Hoechst (Cayman Chemical Company, 15547), UNC3230 (MedChem Express, NJ, USA; HY-110150), remdesivir (AdooQ, Irvine, CA, USA; A17170) and apilimod (MedChem Express, HY-14644). In particular, the purities of remdesivir and apilimod, which were used as antiviral controls, were confirmed to be greater than 95%.
Seo Y., Jang Y., Lee S.G., Rhlee J.H., Kong S., Vo T.T.H., Kim M.H., Lee M.K., Kim B., Hong S.Y., Kim M., Lee J.Y, & Myung K. (2024). A dual inhibitor of PIP5K1C and PIKfyve prevents SARS-CoV-2 entry into cells. Experimental & molecular medicine.
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10

Lysosomal Trafficking and Acidity Assay

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Apilimod (MedChemExpress); WX8 (gift from Juan S. Bonifacino lab) Oregon Green 488-dextran 10,000 mW (OG; Thermo Fisher); chloroquine, BafA1, monensin, and nigericin (Sigma-Aldrich); LysoTracker Blue DND-22 (Invitrogen); Magic Red cathepsin B essay (ImmunoChemistry Technologies); PI(3,5)P2 diC8, PI(4,5)P2 diC8, PI3P diC8 (Echelon Biosciences).
Leray X., Hilton J.K., Nwangwu K., Becerril A., Mikusevic V., Fitzgerald G., Amin A., Weston M.R, & Mindell J.A. (2022). Tonic inhibition of the chloride/proton antiporter ClC-7 by PI(3,5)P2 is crucial for lysosomal pH maintenance. eLife, 11, e74136.
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