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September 5, 2025Open Access

Supplementary Figure S3 from The Discovery and Characterization of HBS-101, a Novel Inhibitor of Midkine, as a Therapeutic Agent for the Treatment of Triple-Negative Breast Cancer

MMMegharani MahajanTexas A&M University – San Antonio ASAlondra L. Rodriguez SanchezThe University of Texas Health Science Center at San Antonio SJSridharan JayamohanThe University of Texas Health Science Center at San Antonio

Key Points

HBS-101 treatment reduces cell viability in triple-negative breast cancer cells, and confirms its potential as a therapeutic agent.
Cell viability assessments using MTT and Cell Titer-Glo assays show significant effects on cancerous cells after HBS-101 treatment.
The analysis employed Western blotting to confirm MDK knockdown and assess the impact of HBS-101 on key signaling pathways.
Midkine inhibition by HBS-101 may alter crucial cellular processes, highlighting its potential for targeted therapy.

Abstract

Supplementary Figure S3: A, Activity of biotin-HBS-101 on cell viability was analyzed by MTT assay (n = 3). B, The sensitivity of both scrambled and MDK-siRNA BT-549 cells to HBS-101 treatment was evaluated by measuring cell viability with the Cell Titer-Glo assay after 72 hours of treatment. C, The Western blot confirms the successful knockdown of MDK. D, Quantification of the fold change in Annexin V-positive MCF-10A cells is presented as a bar graph. E, To evaluate the effect of HBS-101 on MDK-mediated PI3K/AKT signaling, MDA-MB-231 cells were treated with HBS-101 (20 µM, 15 hrs), and the protein expression levels were analyzed by Western blotting.

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