Human PIKFyve Antibody

Detection of Human PIKFyve by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 0.25 µg/mL of Sheep Anti-Human PIKFyve Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7885) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PIKFyve at approximately 240 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of PIKFyve by Western Blot

PIK5-12d depleted PIKfyveprotein in vivo. (A) Studydesign of pharmacodynamic assessment on target engagement of PIKfyvePROTAC degrader in LTL-331R human prostate cancer patient-derivedxenograft model; (B) immunoblotting of PIKfyve, LC3A/B, and GAPDHin the whole cell lysate of human LTL-331R tumors treated with vehicleor various concentrations of PIK5-12d for 4 days; (C)representative images of TUNEL signal of LTL-331R tumors treated withvehicle or 10 mg/kg PIK5-12d for 4 days; and (D) tumorproliferation of LTL-331R tumors treated with vehicle or 15 mg/kg PIK5-12d at 5 days on and 2 days off regimen for 17 days,the p value was determined by two-tailed unpaired t test between vehicle and PIK5-12d groups. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37605297), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of PIKFyve by Flow Cytometry

Genetic or pharmacologic inhibition of PIKfyve upregulates antigen presentation and enhances cancer cell killing by CD8+ T cells. (A) Left: Viability of cancer cells with or without OVA overexpression treated with the indicated agent for 24 h, followed by coculture with activated CD8+ T cells isolated from OT1 mice for 48 h. Right: Viability of control or Pikfyve-null cancer cells with or without OVA overexpression, cocultured with activated CD8+ T cells isolated from OT1 mice for 48 h. (B) Representative images (Left) and quantification (Right) of flow cytometry measuring the proportion of IFN-gamma + and Ki67+ cells in the activated CD8+ T cells cocultured with the indicated cancer cells in (A). (C) Representative images (Left) and quantification (Right) of flow cytometry measuring surface expression of an OVA-peptide (SIINFEKL) bound MHC-I (H-2Kb) in the control or Pikfyve-null cancer cells, treated with or without IFN-gamma at 10 ng/mL for 24 h. Data in A, B, and C were acquired with three independent biological replicates presented as mean ± SD. Statistics were acquired by two-tailed Student’s t test (A and B) or two-way ANOVA (C), with Bonferroni correction. MFI: mean fluorescence intensity. Control: non-targeting single-guide RNA. Pikfyve KO1 and Pikfyve KO2: independent single-guide RNAs depleting Pikfyve. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38011559), licensed under a CC-BY license. Not internally tested by R&D Systems.