Journal: The FASEB Journal

Article Title: Leukemic IL‐17RB signaling regulates leukemic survival and chemoresistance

doi: 10.1096/fj.201900099r

Figure Lengend Snippet: Figure 1. IL-17B and IL-17RB are significantly up-regulated in AML. A, B) Analysis of IL-17B (A) or IL-17RB (B) mRNA expression levels in normal human HSCs (lineage negative, CD34+ CD382 CD90+ CD45RA2), primary AML samples with AML1-ETO fusion protein (n = 39), AML with PML–RAR-a fusion protein (n = 37), AML with inv(16)/t(16;16) (n = 28), and AML with t(11q23)/ MLL (n = 38) using the HemaExplorer database. Data sets from HemaExplorer were first separated into batches according to platform and laboratory. These batches were normalized using robust multiarray average method provided in the affy package in R. Batch effects were corrected using ComBat. C, D) Quantitative RT-PCR assay on the IL-17B (C) or IL-17RB (D) mRNA level in the AML blast–enriched primary BMMCs (n = 12) or normal BMMCs (n = 4). E) Western blotting analysis of IL-17RB and IL-17B (continued on next page)

Article Snippet: Cytokine concentrations were measured using a human IL-17B ELISA kit (Thermo Fisher Scientific) andmouse Il-17b ELISA kit (RayBiotech, Norcross, GA, USA), according to the manufacturers’ instructions.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: The FASEB Journal

Article Title: Leukemic IL‐17RB signaling regulates leukemic survival and chemoresistance

doi: 10.1096/fj.201900099r

Figure Lengend Snippet: Figure 2. Inhibition of IL-17B or IL-17RB leads to decreased viability in AML cells in vitro. A) Western blotting analysis of IL-17RB and IL-17B expression in human AML cell lines. B) Western blotting analysis of the IL-17RB protein level in MOLM-13 and THP- 1 cells transduced with 2 different IL-17RB shRNAs. The NC shRNA was used as a knockdown control. C) Histograms show the relative cell numbers of modified leukemia cells as indicated after culture for 3 d. D) Western blotting analysis of the IL-17B protein level in MOLM-13 and THP-1 cells transduced with 2 different IL-17B shRNAs. The NC shRNA was used as a knockdown (continued on next page)

Article Snippet: Cytokine concentrations were measured using a human IL-17B ELISA kit (Thermo Fisher Scientific) andmouse Il-17b ELISA kit (RayBiotech, Norcross, GA, USA), according to the manufacturers’ instructions.

Techniques: Inhibition, In Vitro, Western Blot, Expressing, Transduction, shRNA, Knockdown, Control

Journal: The FASEB Journal

Article Title: Leukemic IL‐17RB signaling regulates leukemic survival and chemoresistance

doi: 10.1096/fj.201900099r

Figure Lengend Snippet: Figure 3. Silencing of the Il-17b or Il-17rb leads to significant suppression of leukemic progression in vivo. A) Western blotting analysis of Il-17b and Il-17rb expression in murine AML sorted BM c-kit+ leukemia progenitor samples expressing PML–RAR-a, AML1-ETO9a, or MLL-AF9, and a normal sorted BM lineage-negative (Lin2) c-kit+ HSPC sample. B) ChIP-qPCR assay for H3K4me3 (left) and H3K27me3 (right) at the Il-17b or Il-17rb gene locus in the c-kit+ or c-kit2 MLL-AF9+ AML cells as indicated. C) Scheme for the establishment of mouse NC or Il-17b– or Il-17rb–knockdown MLL-AF9+ AML models. D) Western blotting analysis of the Il-17rb protein level in MLL-AF9+ AML cells transduced with 2 different Il-17rb shRNAs. The NC shRNA was used as a knockdown control. E) ELISA analysis showed the Il-17b level in culture medium of MLL-AF9+ AML cells transduced with 2 different Il-17b shRNAs. The NC shRNA was used as a knockdown control. F–K) The recipients were transplanted with 1 3 105

Article Snippet: Cytokine concentrations were measured using a human IL-17B ELISA kit (Thermo Fisher Scientific) andmouse Il-17b ELISA kit (RayBiotech, Norcross, GA, USA), according to the manufacturers’ instructions.

Techniques: In Vivo, Western Blot, Expressing, ChIP-qPCR, Knockdown, Transduction, shRNA, Control, Enzyme-linked Immunosorbent Assay