Additional file 1 of Prolactin receptor signaling induces acquisition of chemoresistance and reduces clonogenicity in acute myeloid leukemia
Creators: name: Cuesta-Casanovas, Laia
name: Delgado-Martínez, Jennifer
name: Cornet-Masana, Josep M.
name: Carbó, José M.
name: Banús-Mulet, Antònia
name: Guijarro, Francesca
name: Esteve, Jordi
name: Risueño, Ruth M.
Publisher: name: figshare
Year: 2023
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: 69999 Biological Sciences not elsewhere classified
subject: Cancer
subject: Hematology

description: Additional file 1: Figure S1. PRLR signaling pathway is differentially expressed in AML in comparison to HSCs and early precursors. A mRNA expression profiles related to PRLR signalingwere selected and hierarchy grouped without supervision, previous normalization by RMA. AML patient samples enrich in LSCs were marked. B PRLRmRNA expression in healthy blood donor cells, ALL, AML, CLL, CML, and MDSfrom the GSR13159 database. **p &lt; 0.01; ***p &lt; 0.001; ****p &lt; 0.0001.. Figure S2. Validation of PRLR-transduced cells and AML murine model. A PRLR surface expression in PRLR wt-transducedand parental controlMonoMac-1, HL-60 and SKM-1 cells by flow cytometry. A representative histogram is shown. B rFLuc-transduced MonoMac-1cells were intravenously injected into adult conditioned NSG mice. At day 4, 6, 8, 11 and 13 mice were treated intraperitoneally with the vehicle, PRL or G129R at 0.2 mg/kg. Engraftment was followed by bioluminescence at day 4, 6, 8, 11, 13 and 14. C PRLR wt-and PRLR short-transducedMonoMac-1cells were validated by qPCRand Western Blotwith GAPDH as loading control and a representative membrane is shown. *p &lt; 0.05; ****p &lt; 0.0001. Figure S3. PRLR signals through Jak2/Stat5. A PRLR-transduced MonoMac-1 cellswere treated with the vehicle, PRLor G129Rat different doses for 2 h and total protein lysates were obtained. Phosphorylated and total Stat5 were analysed by Western Blot; GAPDH was used as loading control. A representative membrane is shown. B CRE and SRE reporters were transfected in HEK293T cells and treated with the vehicle, PRLor G129Rat 500 ng/mL. The luciferase activity is shown in a representative replicate. **p &lt; 0.01. C MonoMac-1and HL-60were treated with a Stat3, Stat5 and Jak2 inhibitors for 72 h at different doses and viability was analysed by flow cytometry. A representative result was shown. Bars represent the mean ± SEM. Figure S4. PRLR confers chemoresistance to AML. A PRLR surface expression analysis in non-refractoryvs. refractoryAML patient samplesby flow cytometry. B Cytarabine-resistantand the parentalcell lines HL-60 and SKM-1were validated by flow cytometry. A representative result is shown. Figure S5. PRLR overexpression does not affect AML proliferation capacity. A Proliferationwas assessed in PRLR-transducedand parental controlHL-60 and SKM-1 cells by flow cytometry. B p16 mRNA expression in PRLR-tranducedand parental controlHL-60 and SKM-1 cells analysed by qPCR. C Cell cycle analysis of PRLR-transduced and parental control HL-60 and SKM-1 analysed by flow cytometry. A representative histogram is shown. Figure S6. A graphical summary of the role of PRLR-PRL signaling in AML.