Chromospheric Activity of HAT-P-11: an Unusually Active Planet-Hosting K Star

Kepler photometry of the hot Neptune host star HAT-P-11 suggests that its spot latitude distribution is comparable to the Sun's near solar maximum. We search for evidence of an activity cycle in the CaII H & K chromospheric emission $S$-index with archival Keck/HIRES spectra and observations from the echelle spectrograph on the ARC 3.5 m Telescope at APO. The chromospheric emission of HAT-P-11 is consistent with a $\gtrsim 10$ year activity cycle, which plateaued near maximum during the Kepler mission. In the cycle that we observed, the star seemed to spend more time near active maximum than minimum. We compare the $\log R^\prime_{HK}$ normalized chromospheric emission index of HAT-P-11 with other stars. HAT-P-11 has unusually strong chromospheric emission compared to planet-hosting stars of similar effective temperature and rotation period, perhaps due to tides raised by its planet.Comment: 16 pages, 8 figures; accepted to the Astrophysical Journa

Additional file 1: of BET Bromodomain inhibition promotes De-repression of TXNIP and activation of ASK1-MAPK pathway in acute myeloid leukemia

Supplementary information. (DOCX 197 kb

Additional file 2: of A loss-of-function genetic screening reveals synergistic targeting of AKT/mTOR and WTN/β-catenin pathways for treatment of AML with high PRL-3 phosphatase

Figure S1. Relative quantification of PTP4A3 (PRL-3) expression level in OCI-AML2 and MOLM-14 cells by qRT-PCR analysis. Figure S2. Representative immunoblot showing the levels of indicated proteins in OCI-AML2 (PRL-3 low) cells with different treatments. Figure S3. In vivo efficacy of VS-5584, ICG-001 single agent and combination treatment in mouse xenograft models transplanted OCI-AML2 cells. (PDF 246 kb

Additional file 1: of A loss-of-function genetic screening reveals synergistic targeting of AKT/mTOR and WTN/β-catenin pathways for treatment of AML with high PRL-3 phosphatase

Table S1. Genes critical for survival of PRL-3 high AML cells revealed by Mission shRNA library screening. (XLS 246 kb

Effects of D9 on histone methylation and transcriptome in AML.

<p>Western blot analysis showing the level of cleaved PARP, EZh2 and a series of histone lysine methylation marks in MOLM-14 (a) and KG-1a cells (b) treated by D9 at indicated concentrations for 48 and 72 hours. (c) Heat map of differential genesets between sensitive and resistant cell lines with D9 treatment. Three sensitive (MOLM-14, MV4-11 and TF-1) and three resistant cell lines (Mono-Mac-1, KG-1a and THP-1) were treated with D9 at 1 or 5 μM for 48 hours. Total RNA was isolated for microarray and SAM analysis. 327 genes were up-regulated and 220 genes were down-regulated upon D9 treatment in sensitive cells relative to resistant cells using 10% false discovery rate (FDR) cut-off. (d) Ingenuity Pathway Analysis (IPA) of differentially up-regulated geneset and down-regulated geneset (dii) showing their strong connections to PI3K/AKT and MEK/ERK signaling pathways.</p

Effects of D9 on AKT and ERK phosphorylation in AML.

<p>(a)Western blot analysis of p-ERK (T202/204), total ERK, p-AKT (S473), total AKT and ACTIN in indicated AML cell lines treated with D9 for 24 hours and 48 hours. (b) Western blot analysis showing the effects of D9 on Bim and Survivin in AML cell lines as in (a). (c) The frozen primary AML patient blasts were recovered for 24 hours and the dead cells were removed using Dead Cell Removal Kit just before D9 treatment. The blasts were treated for 96 hours for cell viability assay and 48 hours for western blot analysis. The diagrams showed the drug response curves of AML patient blasts towards D9 (ci), measurement of EC50 of D9 using cell viability assay (cii) and western blot analysis of PARP, p-ERK (T202/204), total ERK, p-AKT (S473), total AKT and ACTIN in AML patient blasts as well as MOLM-14 with and without D9 treatment (ciii). Each data point in the plots of drug response curves of D9 represents the mean ± SEM of six replicates at each specified concentration of D9, N = 3.</p

Effective anti-leukemia stem cells (LSC) activity of D9 in AML.

<p>(a) Bar graphs showing FACS analysis of the proportion of CD34+CD38- population in TF-1a after the single treatment with D9, SAHA or DAC for 72 hours. (b) Representative FACS histogram profiles of CD34+CD38- cell population in TF-1a cells treated with D9 (100 nM) alone or in combination with either Ara-C (20 nM) or ADR (50 nM). (c) Bar graphs showing the proportion of CD34+CD38- population in TF-1a cells treated as in B. (d) Bar graphs showing the proportion of CD34+CD38- population in TF-1a treated with D9 (20 nM) with or without Ara-C (20 nM) for 14 days. (e). Bar graphs showing the percentage of CD34+CD38- cell population in TF-1a cells treated as indicated. (f) Colony Formation Unit (CFU) assay showing the effects of D9, SAHA or DAC on basal or Ara-C or ADR-induced colony formation capacity of TF-1a cells. The medium and drugs were replenished every 3 to 4 days and the dead cells were removed by Dead Cell Removal Kit. 1 x 10^3 live cells were seeded with semi-solid colony formation medium and incubated for 2 weeks before enumeration. Representative images of the colony formation of TF-1a were shown on (fi) and bar graphs were shown the colony numbers on (fii). Data are mean ± SEM; N = 3; *P < 0.05, **P < 0.01, ***P < 0.001, ns represents no significance, unpaired two tailed t test.</p

In vivo effects of D9 in AML mouse models.

<p>(a) Subcutaneous xenograft tumor growth of MOLM-14 cells in Balb/c mice treated with D9. 5 x 10^6 of MOLM-14 cells were injected into mice subcutaneously (s.c.). D9 at three different doses (30, 60 and 90 mg/kg) was administered through intraperitoneal (i.p.) injection as qd x 5d/w for 3 weeks in these recipients from day 8 to day 23 after transplantation, with vehicle treatment as control. The graphs show the tumor sizes (ai) and body weight change (aii). Data are mean ± SEM, ***P < 0.001, unpaired two tailed t test. (b) Kaplan–Meier curves of NOD/SCID mice treated with D9. 1 x 10^6 of MOLM-14 cells were injected into sublethally irradiated NOD/SCID mice through tail vein. D9 at 60 mg/kg was administered through i.p. injections as q2d/w for 3 weeks in these recipients from day 2 after transplantation, with vehicle treatment as control. The graphs showed the survival rate (bi) and body weight change (bii) during D9 treatment. Data are mean ± SEM, ***P <0.001, log-rank (Mantel-Cox) test conducted.</p

D9 targets cell adhesion-mediated drug resistance (CAM-DR) in AML.

<p>(a) Heat map of 720 genesets induced by both Ara-C and ADR which were suppressed by D9. (b) The diagram showing the top five Bio Functions of commonly upregulated 720 genes by Ara-C and ADR suggested by IPA. (c) The diagram showing the top ten canonical pathways of commonly upregulated 720 genes by Ara-C and ADR suggested by IPA. (d) The diagrams showing Integrins and AKT are highly associated networks of commonly upregulated 720 genes by Ara-C and ADR suggested by IPA. (e) Bar graphs showing the averaged values of 46 probes of Integrin members, 24 probes of Laminins, 8 probes of cytokines and 8 probes of receptors of cytokines extracted from normalized microarray data of CD34+CD38- double-selected TF-1a cells treated as in A. (f) Bar graphs showing qRT-PCR validation of microarray expression data of 15 representative genes as indicated. (g) Representative phase-contrast images of TF-1a cells treated with D9 (100 nM), Ara-C (50 nM) alone or combination. Bar graphs showing transwell migration assay (h), invasion assays (i) and adhesion assay (j) conducted on TF-1a cells treated with D9 (100 nM), SAHA (100 nM) or DAC (100 nM) with or without Ara-C (100 nM). Data are mean ± SEM; N = 3; *P < 0.05, **P < 0.01, ***P < 0.001, ns represents no significance, unpaired two tailed t test.</p

EC50 profile of D9 in human cancer cell lines.

<p>(a) Chemical structure of D9. (b) Bar graphs showing EC50 of D9 measured in a panel of solid (bi) and blood (bii) cancer cell lines using cell viability assay.1 x 10^³ cells of individual cell lines were seeded into 96-well plates in triplicates and D9 at 10 different doses was added 24 hours post cell seeding. Proliferation was measured after 96 hours treatment of D9 using an ATP based cell viability assay. EC50 of D9 was calculated by nonlinear regression (curve fit) using GraphPad PRISM3. Data represent the means of EC50 of D9 measured in three independent experiments, with each experiment run in triplicates.</p