Additional file 1 of Expression of TFRC helps to improve the antineoplastic effect of Ara-C on AML cells through a targeted delivery carrier
Creators: name: Wu, Xinzhou
name: Jiao, Zhouguang
name: Zhang, Junying
name: Li, Feng
name: Li, Yuhua
Publisher: name: figshare
Year: 2023
Subjects: subject: Biophysics
subject: Biochemistry
subject: Physical Sciences not elsewhere classified
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Cancer
subject: Hematology

description: Additional file 1: Fig. S1 Characterization of TFRC gene expression in cells obtained from different sources. a The expression levels of TFRC in bone marrow endothelial progenitor cells isolated from AML patients or healthy donors (GSE197907, n=3). *p&lt;0.05, t test. b The expression levels of TFRC in bone marrow immune cells isolated from AML patients or healthy donors (GSE154109, n=4 and n=8 in healthy donors and AML patients). *p&lt;0.05, t test. c TPM changes in TFRC gene expression in AML cells based on RNA-seq data after treatment with Ara-C (GSE145061, n=3). *p&lt;0.05, t test. d FPKM changes in TFRC gene expression in AML cells based on RNA-seq data after treatment with Ara-C (GSE146592, n=6 on Day 0, n=4 on Day 8, and n=5 on Day 29). *p&lt;0.05, Student‑Newman‑Keuls test. TPM, transcripts per million; FPKM, fragment counts per kilobase of transcript sequence per million base pairs sequenced. Fig. S2 Assessments of intracellular internalization and cytotoxicity of Ara-C@HFn on HL-60 cells blocking with anti-TFRC antibodies. a, b Representative confocal laser scanning microscope images of Ara-C@HFn (Cy5 labeled) endocytosed by normal HL-60 a and anti-TFRC antibody blocked HL-60 b cells (green, lysosome; blue, nucleus; red, Cy5–Ara-C@HFn; scale bar, 5 μm). c Relative viability analysis of normal HL-60 and anti-TFRC antibody blocked HL-60 cells after treatment with different concentrations of Ara-C@HFn for 48 h (n=4). d The kinetics of Ara-C release from Ara-C@HFn in vitro at pH 5.0 and pH 7.0 at 37 ℃ (n=3); e Percentage analysis of the G0/G1, S and G2/M phases of the cell cycle in HL-60 cells treated with free Ara-C or Ara-C@HFn (n=3). f Ratio analysis of the percentage of S phase and G0/G1 phase of the cell cycle in HL-60 cells after treatment with free Ara-C or Ara-C@HFn (n=3). *p&lt;0.05, Student‑Newman‑Keuls test. Fig. S3 Assessments of Ara-C@HFn targeting ability in leukemia cells in vivo and expression characteristics of TFRC in AML cells after Ara-C treatment. a Flow cytometry analysis of Cy5-Ara-C@HFn binding to cells extracted from the peripheral blood, hindlimbs or spleen of leukemia-bearing mice (n=3). b Percentage of hCD45+ cells from HFn+ cells extracted from the peripheral blood, hindlimbs or spleen of leukemia-bearing mice (n=3). c TFRC expression in AML cells (hCD45+ cells) in peripheral blood obtained from mice in the three treatment groups (n=4). Fig. S4 Biosafety evaluation of Ara-C@HFn. a–c WBC, PLT and RBC counts in peripheral blood obtained from mice treated with PBS, free Ara-C or Ara-C@HFn (n=3). *p&lt;0.05; **p&lt;0.01; Student‑Newman‑Keuls test. d The concentration of HGB in peripheral blood obtained from mice treated with PBS, free Ara-C or Ara-C@HFn (n=3). e Histopathological analysis of tissue sections from mice treated with PBS, free Ara-C or Ara-C@HFn (n=3). Fig. S5 Characteristics of TFRC expression in AML patient-derived leukemia cells in various clinical scenarios. a TPM expression of TFRC in AML cells with different expression levels of GPR56 or CD34 based on RNA-seq data (GSE129094, n=7 in the GPR56− CD34− group, n=7 in the GPR56+ CD34- group, n=6 in the GPR56+ CD34+ group). b–d The TFRC levels in AML cells from newly diagnosed and relapsed AML patients b, GSE156008, n=9 in the newly diagnosed group and n=15 in the relapsed group; c, GSE199451, n=11; d, GSE153348, n=6). e, f The TFRC levels in AML cells from the CR group and non-CR group (e), GSE103424, n=18; f GSE164894, n=15 in the CR group and n=9 in the non-CR group). (g) The TFRC levels in AML cells obtained from progressor and non-progressor groups among AML patients administered combined venetoclax and azacitidine treatment (GSE155431, n=6 in the non-progressor group and n=3 in the progressor group). h TPM expression of TFRC in AML cells of non-responder, partial responder and responder groups among AML patients treated with fedratinib (GSE199455, n=3, n=4, and n=8, respectively). TPM transcripts per million, CR complete remission, non-CR noncomplete remission.