EMBO/sd-character-level-ner huggingface dataset:
Kaplan-Meier curves showing mammary tumor-specific survival for the different models. WapCre;Brca1F/F;Trp53F/+;Col1a1invCAG-BE3/+ females injected with Lenti-sgPik3caE545K/sgTrp53Q97*-Myc (n=6) showed a reduced mammary tumor-specific survival compared to animals injected with Lenti-sgTrp53Q97*-Myc (n=5) vectors (76 days after injection vs 101 days after injection, P < 0.05 by Mantel-Cox test). Females injected with Lenti-sgNT-Myc (n=11) did not develop any palpable tumors during the 150 days observation period.
(J) Top chamber: WT BMMs; Bottom chamber: WT or Mavs -/- BMMs. Data shown in (J) are the mean ± SD (n = 3 wells per group) and all data shown are representative of three independent experiments (biological replicates). * p < 0.05, ** p < 0.01 and *** p < 0.001 by Student's t-test (two tailed).
ELISA detection of IFNβ release by mBMDMs treated for 4 h or 24 h with extracellular c-di-AMP at indicated concentrations.
Selected regions from 1H-15N TROSY spectra of 2H,13C,15N-labelled R0RBR parkin bound to unlabelled pUb in the absence (black contours) and presence (blue contours) of one equivalent invisible 2H-labelled UbcH7-Ub. A large number of signals for residues including S145, N235, S255, E399, Q400, W403, E404 and D464 either shift or are absent in spectra with UbcH7-Ub.
A. Schematic representation of zebrafish Endouc protein and its internal deletion mutants.
Quantification of aggregated and total muHTT in the infused striatum of HD mice after 4-week cholesterol infusion by TR-FRET analysis using different antibody pairs. Schematic representation of employed TR-FRET assay (I).
H-K) Representative images showing that Hh overexpression using pan-glial (repo-GAL4) and cortex-glial (NP2222-GAL4) drivers both result in a decrease in NB EdU index, quantified in (J) (n=16, 20 brain lobes) and (K) (n=11, 14 brain lobes), respectively.
(f,g) Nutrients and pHi affect levels of ARL8 and KIF2 in lysosomal fractions. Protein levels and their quantification in total cellular lysates or in isolated lysosomal fractions from HeLa cells subjected to 1 h nutrient deprivation-recovery (f) or to 1 h changes of pHi in full tissue-culture medium containing nigericin (g) are shown.
BMDMs were costimulated with TNF (100 ng/ml) and TAK1i (125 nM) for 4 h and mixed supernatant and extracts were analysed by immunoblot. Where indicated, cells were treated with the inhibitors MCC950 (10 μM), GSK'872 (1 μM), 20-30 min prior to cell stimulation. KCl (50 mM) was added together with TNF and TAK1i. To induce necroptosis, BMDMs were primed for 3 h with ultrapure E. coli K12 LPS (100 ng/ml) and Q-VD-OPh (10 μM) was added at the last 20-30 min of priming and stimulated with SM (500 nM) for 4 h.
(A, B) Co-immunoprecipitation of ectopically expressed Myc-CDC20 171-499 fragment (containing WD40 domain) (A) rather than Myc-CDC20 1-170 fragment (lacking WD40 domain) (B) with endogenous p65 in HEK293T cells.
(d) Bax+/+Bak+/+ (WT) and Bax−/−Bak−/− (DKO) MEFs were cultured in EBSS in the presence of 50 μM z-VAD-fmk or control DMSO for 0, 6 or 12 h.
Quantification of mitotic cells in wild-type animals fed as (A) and left untreated or treate with 1200 J/m2 UV Data information: Data are presented as mean ± SD. For each panel N=2, 8-10 worms per replicate and condition
Q As in (I), but for SUN1 staining. Arrowheads in (Q) indicate the position of altered telomeres. Data information: Sex bivalents (XY) are indicated. Scale bar, 10 μm.
(h) Immunoblot of LC3 and p62 in control (n = 10) and CF (n = 10) human nasal mucosa. (i) Number of LC3-positive dots (per 104 mm−2 of mucosa) in control (n = 10) and CF (n = 10) human nasal mucosa. Means ± s.d. Asterisk, P 0.01 versus controls; ANOVA.
D. Using high power, high resolution laser scanning confocal microscopy, no mouse LAG3 signal could be detected in murine primary neuronal cultures (Auto-mLAG3 transduced, DOX OFF and non-transduced neurons) using anti-mouse LAG3 antibody (MABF954) whereas LAG3 was clearly detected in primary neurons induced to express mLAG3 (Auto-mLAG3 transduced, DOX ON). Scale bars 20 µm.
D Enhancer activity was compared between literature‐based ARE and Exo‐defined ARBE in TMPRSS2 locus by reporter gene assay.
C) Scatterplot indicating the BrainSpan cell cycle module trajectory across development (vertical line indicates birth; time-points to the left of the line are fetal time-points, while time-points to the right of the line are postnatal time points; best-fit curve indicates a 4th order polynomial fit).
Real‐time PCR for SFB in luminal, mucosal, and fecal samples of multiple ANA‐positive and ANA‐negative Ltbr−/− mice. Analysis with ANOVA, P = 0.02 for differences between groups, n = 4 mice per group. Data represent means ± s.e.m.
c) DAVID functional annotation clustering analysis showing the most significantly overrepresented GO terms in age-downregulated miR-29 targets.
Western blot analysis of (E) Fmrp expression (S n=11, SD n=10) in the prefrontal cortex of sleeping and sleep deprived mice. Student's t-test *p < 0.05, **p < 0.01, ***p < 0.001.
(B) Co-immunoprecipitation and immunoblot analysis of extracts of NS1-inducible THP-1 cells, which expressed Flag-tagged NS1, treated with or without doxycycline (Dox) (100 ng/ml).
H 293T cells were transfected with plasmid encoding Flag-Beclin-1 and treated with CHX (100 μg/ml) alone or with MG132 (10 μM), 3MA (10 mM), CQ (50 μM) as combination for 6 h or transfected with the HA-USP19 plasmid. The cell lysates were analyzed by immunoblot.
B Latency to enter dark compartment for Gadd45a-WT (white bars, n=11) and Gadd45a-KO (blue bars, n=11). A significant reduction was found in the Gadd45a-KO at 24 hours and 7 days after the training. Values shown are mean ± SEM; 2-Way ANOVA and Bonferroni post-hoc test: * = p<0.05.
F) Plaque assays were performed similarly as described for (A) with RH (n=5 for plaque number and n=3 for plaque area) and RH+GRA15II (n=5 for plaque number and n=3 for plaque area).
B CD81 facilitates CD19 maturation. CD81 and CD19 (300 ng DNA) were transfected into the CD81-knockout 293T cells. Anti-FLAG immunoblot was performed for CD19 with a C-FLAG tag. The deglycosylation was performed by digesting the cell lysate with PNGase F or Endo H, following the manufacturers' instructions. The deglycosylation pattern of CD19 species (p: precursor, m and m': two mature forms) differs because the glycosylation content of these species is different
A. The structure of the imp and hcp operons and vgrG2 operon in A. tumefaciens C58 [10]. The arrows represent coding sequencing, with arrowheads depicting the direction of expression. The wider arrows represent genes in the T6SS associated operons. The vgrG2 operon is located distal to imp operon and hcp operon. The vgrG and toxin-immunity gene pairs are highlighted in colors.
(D) Lesion progression in BALB/c mice inoculated with 5×105 stationary phase promastigotes. Values shown are the means ± SD from 5 mice. *, infection level of Δatg5 differed significantly from WT (p<0.01) and Δatg5::ATG5 (p<0.05).
(A) Domain structure of human UBQLN1 (Interpro: http://www.ebi.ac.uk/interpro/).
E Induction of Tregs with PGE2 analog (DINP) (5µM) and/or recombinant human IL-10 (rhIL-10) (10ng/ml) as compared to CLys.
(A-A''') Immunostaining of discs expressing ptc>RFP with antibodies to Rpk and ATPα showing elevated levels of both proteins anterior to the compartment boundary. White arrowheads indicate approximate position of the A-P compartment boundary.
Forebrain slices from S1928A KI mice were treated with vehicle (lanes 1,5) or 10 M ISO for 5 min, followed, if indicated, by 1, 3, or 20 min wash out of ISO (lanes 3-7) and a second application of ISO for 5 min (lane 6), before solubilization and ultracentrifugation.(A) α11.2 and GluA1 were concurrently IPed from same samples by simultaneous addition of anti-α11.2 and -GluA1 antibodies before probing and stripping/re-probing upper part of IB for pS1928, pS1700, and total α11.2 and middle part for pS845, pS831, and total GluA1. In S1928A KI mice, S1700 re-phosphorylation after a 3 min wash out of ISO was not blunted (lane 6) in contrast to WT mice but rather augmented (compare to lane 5).(B-E) For quantification, pS1928 (virtually absent) and pS1700 IB signals were normalized to total α11.2 and pS845 and pS831 signals to total GluA1 (**p<0.01, ***p<0.001, ANOVA).
(K) Mcp3 lacking either of the two transmembrane domains is unstable. Crude mitochondria were obtained from wild-type cells containing an empty plasmid (Ø) or mcp3Δ cells (Δ) transformed with plasmid encoding Mcp3 or Mcp3 lacking TMD1 or TMD2 (MCP3, ΔTMD1, ΔTMD2). Samples were analysed by SDS-PAGE and immunodecoration with antibodies against Mcp3 and Tom20 as loading control.
(E, F) Mean±SEM fluorescence intensities of the indicated Far1(1-150)-GFP mutants over the cell cycle.
(B) Scheme representing the study design. Vectors were injected in C57Bl/6 mice (n=8) at a dose of 1E+10 vg/mouse. Red symbols represent timing of blood collection.
F. Q-PCR analysis of miR-23a, miR-27a and miR-24 expression in ESCs (JM8A3) and a variety of C57BL/6J adult tissues. Data are shown as mean + s.d. (n=3).
(a) Overview of enteric fever cohorts used in this study (T1: Typhoid CHIM study 1; T2: Typhoid CHIM study 2; P1: Paratyphoid CHIM; 03NP: Nepali cohort. ST: S. Typhi; SPT: S. Paratyphi A; sEF: suspected Enteric Fever; D0: day of challenge which represents the control samples in the Oxford CHIM; CTRL: endemic community controls; nD7: day 7 after challenge in participants who stayed well in the CHIM; BC+: Blood-culture positive; BC-: Blood-culture negative; Dx: Diagnosis).
Immunoblot of 6E10 anti-APP immunopurifications from the conditioned media of CHO cells or CHO-derived APP751-expressing (7WD10) cells overexpressing ERdj3WT, ERdj3H53Q, and BiP as indicated. Media inputs (1:100) are shown as a control. Lysates from these cells are also shown to confirm the increase of BiP afforded by overexpression.
E Read coverage of SMG5 from WT HEK293 and HeLa RNA-seq data with or without induced UPF3A overexpression shown as Integrative Genomics Viewer (IGV) snapshot. Differential gene expression (from DESeq2) is indicated as log2 fold change (log2FC) on the right. Schematic representation of the protein coding transcript below.
RNAs from isolated nuclear and cytoplasmic fractionation were harvested after RNA immunoprecipitation (RIP) of RNA-protein complexes using either anti-QKI antibody or control IgG. And RIP-RT-PCR showing the PGC1α and UCP1 transcripts majorly interacted with endogenous QKI in the nucleus. (n=3 biological replicates)
(F) Analysis of NRF2 ubiquitination in absence and presence of TRIM16 deletion variants by Co-IP assays using transiently transfected plasmid constructs as indicated.
Rectal temperature was measured when mice in (A) was challenged to 4˚C. n=5 mice each group. Data information: Data represent means±SEM. *P<0.05; **P<0.01; ***P<0.001; unpaired two-tailed Student's t test.
B to D. Determination of anti-HBV efficacy and safety by measuring HBsAg in the serum of treated mice (B).
MEF2-lacZ reporter mice (BALB/c-background, 6-12 weeks old) were treated with 7 mg/kg lipopolysaccharide from Escherichia coli (O111:B4) or saline intraperitoneally and were sacrificed after 24 hours B PGE2 was quantified after mechanical homogenization of deeply frozen hearts using nano-liquid chromatography tandem mass spectrometry Data information: values are mean±s.e.m. The exact n values are shown in the bottom of the bar graphs. *,P<0.05, Welch's two-tailed unpaired t-test was used for statistical analysis, n.s.=not significant P values PGE2, P=0.0016 (B
Immunohistochemistry staining for the Paneth cell marker LYZ1 in Mex3a KO and WT intestinal organoid sections at day 2 of culture. Scale bars, 50 μm.
(C) Bacterial counts recovered from the blood and brain in wild type mice (n = 5) and CysLT2-/- mice (n = 5) infected with strain K79 for 1h.
(d) Rheb suppresses Ulk1 Ser 317 and Ser 777 phosphorylation in a manner dependent on mTORC1. HA-Ulk1, AMPKα kinase-dead mutant (DN), and Myc-Rheb were co-transfected into HEK293 cells as indicated. The cells were incubated with glucose-free medium (-Glu, glucose), in which either 20 μM compound C (C.C.) or 50 nM Rapamycin (Rapa) was added. Total cell lysates were probed with antibodies against Ulk1 phosphorylated at Ser 317, Ser 777, Ser 757, and HA, as indicated.
(A) Significantly deregulated canonical pathways in GL261 Rheb1fl/fl versus Rheb1Δ/Δ TAM-MG, as identified by the IPA software from differentially regulated genes. Size of bubbles is indicative of ratio of differentially regulated genes. The x-axis represents -log10(p-value) with a threshold of p≤0.05 applied. On the right, number of up and down regulated genes in each identified pathway is indicated in blue and red respectively.
D. HNF1A mRNA levels differed in HNF1A LoF and control groups (Kruskal-Wallis, P<0.01), despite considerable variability and overlap between groups. E. KDM6A mRNA levels were downregulated in HNF1A LoF tumors (Kruskal-Wallis, P<0.001).Data information: Box plots in (D and E) show HNF1A and KDM6A expression in HNF1A LoF tumors (n=26) and Control 1 (n=39) and Control 2 (n=57) tumors. The horizontal central line marks the median. Box limits indicate the first and third quartiles and whiskers extend to highest and lowest data points within 1.5 x interquartile range outside box limits.
(A) The expression of dBI‐1 was knocked down in Drosophila melanogaster. Then, relative expression levels of dBI‐1 mRNA were monitored by semiquantitative PCR. Actin levels were monitored as control.
G A transcriptional AtMYB64 reporter (AtMYB64-NYG) is specifically expressed in all four cell types of the A. thaliana embryo sac Scale bar: 25 µm. Green, YFP fluorescence; white, cell walls.
WT MEF. Scale bar: 10 μm. D Ectopic expression of FAM134BLIR in WT MEF inhibits ATZ delivery to endolysosomes. Scale bar: 10 μm.
A. Transmission EM demonstrates the absence of synaptic ribbons in presynaptic terminals of photoreceptorribbon synapses in RBEKO/KOmice, while synaptic ribbons are readily visible in the terminals of RBEWT/WTmice. Asterisks in A2 and A3 denote photoreceptor active zones of RBEKO/KOmice in which the synaptic ribbon is absent. Except for the absence of synaptic ribbons, the ultrastructural appearance of the presynaptic terminals is comparable to that of RBEWT/WTmice. Abbreviations: sr, synaptic ribbon; pre, presynaptic; post, postsynaptic. Scale bars: 500 nm.
The normal MCF7 (left) and T47D (right) cells were cultured in the media collected from the corresponding cells 48 hours after siRNA-mediated gene knock-down. Expressions of the ISGs in these normal cells were measured with qPCR. Each sample has 3 replicates. Error bars represent mean ± SD.
F. Relative expression of antioxidant genes in NTG and TG mice. N=6 mice in each group. Two-tailed unpaired T-test was performed. All data are expressed as mean ± SEM. N.S. = not signigicant.
JNK signaling is activated during a distinct phase of EMT and becomes a critical determinant of mesenchymal identity. (A) Heatmap showing standardized expression (Z-scores) of genes differentially expressed upon TGF-β-induced EMT in NMuMG cells.
Mice were pre-treated with LicoB or MCC950 for 1 h, then i.p. injected with LPS (20 mg/kg) and treated for 3 h. Quantification of peritoneal exudate cells (PECs) (F) and macrophages (F4/80+ cells) (G) using flow cytometry (n=8 mice per group).
(A-B) Volcano plots visualizing the differentially expressed genes in ARLmon/Y (A) and AR-/Y (B) testes compared to testes from WT males. The dotted horizontal lines represent a q-value of 0.05, while the dotted vertical lines represent a FC of 1.5. Genes that are downregulated compared to WT are shown using blue dots, while red dots represent the upregulated genes. Hsd17b3 and Star are indicated as yellow and green stars, respectively.
(A) Nucleotide sequence alignment of putative stem-loop structures in the C. elegans sams-3, sams-4 and sams-5 genes together with six hairpin structures in 3'UTR of the human MAT2A gene. Conserved residues are shaded in black. Loop, transition and stem regions of the MAT2A hairpins (Doxtader et al, 2018) are boxed in magenta, green and grey, respectively. An arrowhead indicates adenine bases in the MAT2A hairpins that are specifically modified into m6A by METTL16 in vitro (Shima et al, 2017).
D,E Graph presentation of effects of H2A mutations (D) and H2A.B mutations (E) on nucleosome resistance to MNase digestion. MNase digestion efficiency for each sample is calculated from quantitation of the digested DNA against total DNA. All experiments are repeated twice. Data are mean ± SD, n = 2.
(B) Vacuoles were isolated from the indicated yeast strains and 30 µg of proteins analyzed by SDS-PAGE and Western blotting.
Scatter plot and correlation coefficient (r) of ASK1 mRNA expression and BMI Data were collected in lean subjects (n=14), obese subjects (n=23) and obese subjects with type 2 diabetes (n=19). Values are expressed as mean ± SEM (D).
Figure 4e. Rescued loading of CDC6 and CDT1 on chromatin after removal of CDK12 inhibitor. Western blot analyses of chromatin fractions of serum starved AS CDK12 HCT116 cells treated with 3-MB-PP1 for 6 or 9 h or with the inhibitor washed off after 1 h of treatment. CTRL corresponds to cells not treated with the inhibitor at the time of the serum addition. All cells were harvested either 6 or 9 h after the serum addition. Histone H2A serves as a loading control of chromatin fractions and studied DNA replication factors are indicated. A representative image of three replicates is shown.
G-H. Scatter plot showing a significant positive correlation between the indicated features in DLD-1 cells (n = 1). The lines represent linear regression with 95% confidence band. Data from chr 13, 14, 21, 22, Y and SN were excluded from the analysis.
Representative images of CRTH2 subcellular localization in NIH 3T3 cells in response to TGF-β1 (10 ng/mL) treatment. Red arrows indicate CRTH2 localized in the ER and white arrows indicate CRTH2 localized in the plasma membrane. Green, GFP-CRTH2; blue, DAPI; red, Calnexin; Scale bar, 20 μm.
A AMPHIPASEEK prediction result for Bacillus subtilis Noc. The red "A"s indicate a putative amphipathic helical region.
Levels of VPS16 transcripts in fibroblasts by qPCR. The probe of the exon 22-24 PCR spans the misspliced intron-exon border. Data was normalized to levels of POLR2A using the ∆∆Ct method.
A) THP-1 monocytes were genetically engineered to knockout RIG-I, MDA5, or LGP2 using CRISPR/Cas9. Cells were differentiated towards macrophage-like cells using PMA and treated with recombinant type I IFN to upregulate RLR expression. Correct gene editing and intact type I IFN responsiveness was validated by SDS-PAGE and immunoblotting using the indicated antibodies (n=3). *, non-specific band.
(B) Localization analysis of BAG3 and SODG85R in surviving spinal cord motor neurons fromSODG85Rmice at the time of disease symptom onset. Arrows indicate perinuclear aggresomes containing BAG3 and SODG85R. Scale bar, 10 μm. Magnific., magnification; SOD, superoxide dismutase; SODWT, wild‐type SOD.
B. Limited proteolysis of a purified preparation of the Nse5/Nse6 complex by trypsin. Samples taken at different time points were analyzed by SDS-Page and Coomassie staining. Selected stable fragments identified by mass spectrometry are indicated.
A Heatmaps of H3K4me1, H3K27ac, H3K27me3 and H3K9me3 histone modifications in FL erythroblasts around SclMES binding sites in cluster L and cluster R show no gain of common repressive histone marks in cluster L.
[A] Clustering of DA-regulated immunity-related genes in Arabidopsis with sufficient Pi, as revealed by RNAseq analysis.
(D) Islet graft survival of five groups of mice was assessed by monitoring blood glucose and calculated using the Kaplan-Meier method. Cumulative data from two independent experiments are shown. Statistical analysis was performed with a log-rank test. *P<0.05, **P<0.01.
B. Top. Representative immunoblots from GluA2 coimmunoprecipitations probed for Sec23 and GluA2 after no treatment, 10 min DHPG, and and cycloheximide followed by 10 min DHPG. Bottom. Quantification of Sec23 normalized to GluA2 under each of these conditions, For 10 min DHPG, n = 8; for 10 min DHPG + CHX, n = 5, p < 0.05, ANOVA.
(I) Lysates from MCF7 cells transfected with the indicated ubiquitin-HA constructs were subjected to anti-CIP2A immunopurification followed by immunoblotting with antibodies against CIP2A and HA.
E Disruption of the Nap1-histone interaction by H2A mutation. Bottom panel: Inputs of the coexpressed proteins (soluble extract) and top panel the material bound to the glutathione affinity column were analyzed by SDS-PAGE and Coomassie staining.
(D) The ratio of weight of iWAT, eWAT and BAT to body weight in 16-week-old Ftoflox/flox and FtoAKO mice fed a HFD. Data information: The data were presented as the mean ± SD (n = 6). Statistical analyses were performed using Student t‐test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
Multiple sequence alignment of 3 ORF's (KMP91697.1, KMP91698.1 and KMP91699.1) corresponding to DUF3992 containing proteins. Sequence motifs corresponding or similar to that deduced from the EM potential map are shaded in cyan. Secondary structure and structural elements as determined from the built model (see Fig 2) are shown schematically above the sequences (Ntc: N-terminal connecter; arrows correspond to β-strands, labelled as in Fig 2). Contiguous areas of >50% similarity are boxed blue.
His-Activin A-Atto647N was imaged free in solution with no cells (A) or with parental HEK293T cells (B), ACVR2A/B dKO cells (C), parental HEK293T cells pre-incubated with 1.5 μg/mL of rhACVR2A Fc and 15 μg/mL of rhACVR2B Fc (D), HOM1 cells Number and brightness (N&B) analysis was performed on raster-scanned images, and a representative experiment is shown. Panels i, Brightness (B) vs intensity 2D plots. Red rectangular box, pixels that contribute to brightness of free, dimeric His-Activin A-Atto647N in solution. Green rectangular box, pixels that contribute to His-Activin A-Atto647N clustered into putative tetramers. Blue and pink rectangular boxes, pixels that contribute to His-Activin A-Atto647N in higher order clusters. Panels ii, Selection map of the image captured reconstructed with the pixels in red, green, blue and pink rectangular boxes shown in the upper panels (i). Panels iii, B histograms, frequency distribution of the number of pixels across various brightness of His-Activin A-Atto647N of the selected image. Bav indicates the average brightness for each image. Panels iv, cumulative B histograms from a representative experiment with 4 independent regions of interest.
Generation of isogenic mESC lines expressing WT and arginine methylation-deficient mutant (RK) METTL14. Mettl14 KO mESCs were transfected with Flag-tagged WT or RK mutant METTL14 using a lentivirus expression system. The expression of METTL14 and METTL3 in these cells was detected by Western blot analysis using anti-METTL14 and anti-METTL3 antibodies. ACTIN was used as a loading control.
(A) Representative confocal images of mitochondrial morphology in scrambled (Scr) and PERK KD Mfn2 KO cells stained with MitoTracker Green.
Proliferating and senescent IMR90 and HUVECs assayed for β-galactosidase activity (left), cell cycle profiling via FACS (middle) and proliferation via MTT assays (right).
Fluorescence microscopy of GFP-TDGwt GFP-TDGpip in mESCs. Scale bar: 100µM. Quantitation of fluorescence signals of GFP-TDGwt and GFP-TDGpip in mESCs. median with interquartile range. >470 cells were analysed of each GFPTDGwt and GFPTDGpip ESCs.
A mRNA expression in FACS-isolated Lin(Ter119/CD31/CD45)-Sca1+ progenitor cells from female mouse subcutaneous fat, differentiated in the presence of 1 µM cPGI2 or vehicle for the indicated time, as determined by expression profiling (n=3, E-MTAB-3693) ****P=3*10-6 (Day 2), ****P=4*10-7 (Day 4), ****P=1*10-6 (Day 6) in 2x2 ANOVA with Bonferroni posttests (cPGI2 vs. vehicle)