Additional file 1 of Bone marrow-targetable Green Tea Catechin-Based Micellar Nanocomplex for synergistic therapy of Acute myeloid leukemia
Creators: name: Bae, Ki Hyun
name: Lai, Fritz
name: Mong, Jamie
name: Niibori-Nambu, Akiko
name: Chan, Kiat Hwa
name: Her, Zhisheng
name: Osato, Motomi
name: Tan, Min-Han
name: Chen, Qingfeng
name: Kurisawa, Motoichi
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Ecology
subject: Cancer
subject: Hematology

description: Additional file 1: Figure S1. Representative TEM images of (A) Sora-MNC-2 and (B) Sora-MNC-3. Figure S2. Zeta potential distribution profiles of (A) Sora-MNC formulations and (B) HA-EGCG, HA and sorafenib. HA-EGCG (10 mg mL−1) and HA (10 mg mL−1) were dissolved in deionized water. Sorafenib was first dissolved in methanol at 1 mg mL−1 and then 10-fold diluted with water for the zeta potential measurement. Figure S3. Stability of HA-EGCG micelles in 10% (v/v) fetal bovine serum over 5 days. Mean ± SD (n = 3). Figure S4. Anti-leukemic activity of Sora-MNC-1, HA-EGCG and EGCG on (A) MOLM-14 and (B) MV-4-11 cells as a function of EGCG unit concentration. Mean ± SD (n = 4). Figure S5. Median effect plots showing eradication of (A) MOLM-14 and (B) MV-4-11 cells treated with free sorafenib, HA-EGCG or their combination (Sora-MNC-1). Cytotoxicity data were plotted using the linearized median effect equation, log(fa/fu) = m log(D) – m log (Dm), where fa is the fraction of killed cells, fu is the fraction of survived cells, D is the dose applied, Dm is the median effective dose. The resultant plot gave the slope of m and the y-axis intercept of – m log (Dm). Figure S6. Hemolysis assay. (A) Representative photograph of mouse red blood cells treated with PBS, Triton X-100 (1%), or Sora-MNC-1 at various sorafenib doses for 1 h at 37 °C. (B) Effect of different treatments on the absorbance at 576 nm (an indicator of hemoglobin leakage from red blood cells). Figure S7. Flow cytometric detection of CD44 in MOLM-14, MV-4-11 and bone marrow stromal cells labeled without (left panel) or with FITC-tagged anti-CD44 antibody (right panel). Figure S8. (A) Z-average sizes and zeta potential values of Sora-MNC, DyLight 488-labeled MNC and DyLight 800-labeled MNC. (B) Fluorescence emission spectra (λex=400 nm) of DyLight 488 dye (0.312 µg mL−1) and DyLight 488-labeled MNC (Sorafenib concentration=0.8 µg mL−1). (C) Fluorescence emission spectra (λex=750 nm) of DyLight 800 dye (2.5 µg mL−1) and DyLight 800-labeled MNC (Sorafenib concentration=4 µg mL−1). Figure S9. KEGG map of differentially expressed genes (DEGs) involved in the tricarboxylic acid (TCA) cycle. The gradient color scale indicates upregulation (red) and downregulation (green) of gene expression calculated as log2 fold change. The names of DEGs selected for qPCR analysis are highlighted in blue. FH: Fumarate hydratase, SDHA: succinate dehydrogenase, OGDH: 2-oxoglutarate dehydrogenase, IDH1: isocitrate dehydrogenase 1. Figure S10. KEGG map of DEGs involved in the aminoacyl-tRNA biosynthesis. The gradient color scale indicates upregulation (red) and downregulation (green) of gene expression calculated as log2 fold change. The names of DEGs selected for qPCR analysis are highlighted in blue. LARS: leucyl-tRNA synthetase, FARSA: phenylalanyl-tRNA synthetase. Figure S11. Low-magnification images of H&amp;E and immunohistochemical staining of the bone marrow specimen. AML cells were identified with anti-hCD45 antibody (brown area labeled “L”). Scale bars, 200 μm. The red boxes indicate the areas of the higher magnification images included in Figure 6F. Figure S12. (A) Albumin:globulin ratio and (B) blood urea nitrogen level in the experimental groups. Mean ± SD (n = 3); *P &lt; 0.05; n.s.=nonsignificant. Figure S13. (A) Potassium and (B) sodium ion level in the experimental groups. Mean ± SD (n = 3); n.s.=nonsignificant. Table S1. Characteristics of Sora-MNC selected for in vitro studies. Table S2. Sequence of the primers used for qPCR analysis. Table S3. Pharmacokinetic parameters of Sora-MNC and free sorafenib.