EMBO/sd-character-level-ner huggingface dataset:
(C) Kinetic characterization of CK1α inhibition by MDMX. Reaction velocity as a function of β-catenin peptide substrate and increasing MDMX concentrations at 100 µM ATP and 2 nM CK1α. Data were fitted to the Michaelis-Menten equation. The resulting parameter values are shown in the table.
(C) Flow cytometry analysis of cell surface expression of heparan sulfates (HS) in control transduced cells or upon CRISPR/Cas9-mediated EXT1 KO (EXT1-/-).
Structural overlay of the apo (violet) and Ub-bound (pink) wMelOTU structures highlighting the repositioning of the His-loop to accommodate entry of the Ub C-terminus, as well as ordering of two regions in the S1 site that form an embrace around Ub.
F Relative luciferase activities of IFNAR1-3' UTR or IRF3-5' UTR with wild-type (WT) m6A sites in 293T cells that were transfected with poly (I:C)-LMW or infected with VSV-eGFP for 24 hrs. Renilla luciferase activity was measured and normalized to firefly luciferase activity.
(A) Accumulation of GFP-LC3 labeled autophagosomes in Meth- or Clq-treated dendritic cells at 2 h and 24 h. Fisher's exact test P values versus control (n =300-400 cells, mean ± SEM, *p < 0.05, ***p < 0.0001).
(E) Histologic severity scores of COPS8fl/fl mice and COPS8ΔIEC after fecal transplantation. Data represents the histological score of mice analyzed on day 160. Data are represented as mean ± SEM from seven biological replicates per genotype. **P<0.01 using one-way ANOVA.
(E) Cell viability analysis of huDR5 and Chi-G4S-DR5 stable 4T1 cells with indicated human DR5 agonists lexatumumab and KMTR2 (n=3).
Interaction between MARCH2 and NEMO in response to viral infection. Raw264.7 cells (F) were infected with NDV-GFP in a time-dependent manner in the presence of MG132 (proteasome inhibitor, 10 μM). Cell lysates were subjected to immunoprecipitation with an anti-NEMO antibody, followed by immunoblotting with an anti-MARCH2 antibody.
(D, E) Experiments conducted as in (A), but in cells arrested in G1 with alfa-factor (D) or in metaphase with nocodazole (E) for 2 h at 26oC and for one additional hour at 26oC or 35oC.
A. A schematic representation of the three VcdRP and VcdR/P variants used for the transcriptome analysis. Left to right: first, the native VcdRP variant which harbors both the small protein as well as the base-pairing region. Next, a STOP codon was introduced in the 3rd codon of vcdP ORF as a readout for VcdR devoid of VcdP. Finally, the sequence of the VcdP codons was scrambled such that the correct small protein is translated but the nucleotide sequence of the scrambled vcdP variant is significantly altered when compared to native vcdRP. This variant also carries an artificial 5'UTR and a terminator. The arrows indicate the TSS.
(E) Western blot analysis of larval brain extracts of control (ctrl; yw) and pr-set7716/Df at 24h ALH. Blots were probed with anti-Cdk1 antibody and anti-GAPDH1 antibody (loading control).
Daily body weight measured during 7 days after i.c.v. injection of vehicle or AβOs (n = 7 animals/group; data are representative of two independent experiments with similar results).
A Numbers of Paneth cells in the ileum of wildtype and Vil-Ogt KO mice (n = 6, P = 0.0048)
(b) HeLa cells were co-transfected with ULK1-Myc alone or in combination with wild-type ubiquitin or its mutant (K48R or K63R) HA-tagged constructs. Protein extracts were immunoprecipitated using an anti-Myc antibody. Purified complexes and total extracts were analysed by western blotting using anti-Myc and anti-HA antibodies.
H-J. Whole cerebellum lysates of P6, P9 and P12 mice were subjected to immunoblotting with anti-pAtoh1 S309, Atoh1, Ccnd1, Prox1, β-actin. (I) The protein levels of CCND1 and PROX1 normalized by β-ACTIN. (J) Quantification of p-
B Smurf gut barrier assay in anesthetised, live larvae. Left, control larvae, right, larvae overexpressing Atg16B in the gut enterocytes under control of the NP1-Gal4 driver. While the distribution of food in the gut of Atg16B-expressing larvae was different from controls, there was no leakage of dye from the gut in any of the animals in three independent experiments (each with n=30 larvae for each genotype).
(H) The overexpression (OE) of WT UFD-2 or SPR-5 in the C. elegans nervous system blocks the protection conferred by the double mutation ufd-2(tm1380);spr-5(by134) (n = 16). Data represent means ± SEM. The numerical data used to make this figure can be found in S1 Data.
(F) HeLa cells were transfected with control plasmid (pMKO)/GFP-PX or pMKO-BimshRNA/GFP-PX (3:1). The percentages of cells with GFP-vesicles were assessed. Data are shown as mean ± SD. ∗∗p < 0.01.
A-C. While the level of overexpressed GFP-Snc1-PEM is increased in atg9∆ and atg2∆, but not atg18∆, mutant cells, it does not accumulate in aberrant structures and does not induce UPR. Wild type (WT), atg9∆, atg18∆, and atg2∆ mutant cells overexpressing GFP-Snc1-PEM were analyzed as described for Fig 1A-1C, respectively. The tested phenotypes: the level of GFP-Snc1-PEM protein (A), accumulation of GFP-Snc1-PEM in aberrant structures (B), and induction of the UPR response (C,atg1∆ is shown as a positive control).
Western blots for detection of TCF7, LEF1, TCF7L1, TCF7L2 and β-catenin (B") proteins in wild-type (WT) and knockout (K.O.)
B, C. HeLa cells were stably transfected with the indicated vectors, treated with MG132 and immunostained with the indicated antibodies. Scale bar=10 µm.
(H) The frequency of the different Beclin‐1 phenotypes depended on the efficiency of Beclin‐1 knockdown. Mitotic progression in cells treated with increasing concentrations of Beclin‐1 siRNA was followed by video‐microscopy. Left panel: western blot of Beclin‐1 upon RNAi treatment; right panel: quantification of the appearance of the three different mitotic progression patterns, that is, prolonged mitosis with normal anaphase (as C), with lagging chromatids (as D) and mitotic exit without clear anaphase (as E) upon increased Beclin‐1 knockdown experiments. NEB, nuclear envelop breakdown; RNAi, RNA interference; siRNA, small interfering RNA; YFP, yellow fluorescent protein.
(E) Snap-shot images show the effect of CCB02 on microtubule dynamics in two-centrosome containing MCF10A cells. The microtubule was stained with SiR-tubulin and +end tip of microtubule was marked with EB3-EGFP. A 5µM CCB02 did not alter localization and dynamics of EB3 at the microtubules indicating that the CCB02 does not affect microtubule dynamic At least, 50 cells were analyzed for each condition. Scale bar, 2µm
(D-E) WT and Irf3‑/‑Irf7‑/‑ BMDCs were mixed at different ratios for a total number of 2x105 cells/cm2 and stimulated with poly(I:C) or left unstimulated. (D) Representative dot plots showing CD86 and CD40 expression on WT (blue dots) and Irf3‑/‑Irf7‑/‑ (orange dots) BMDCs. (E) Flow cytometric quantification of CD86 and CD40 expression on WT (blue dots) and Irf3‑/‑Irf7‑/‑ (orange dots) BMDCs expressed as a fold change in gMFI over unstimulated controls.
D. Analysis of IL-1R1 expression by Th17 cells isolated from CNS of EAE diseased mice, shown in (B). Data are representative FACS histogram overlays of cells isolated from (grey) wild type mice treated with PBS, (black) wild type, (blue) IL-1R1ΔT and (red) IL-1R1-/-mice treated with PC61, gated on VD-TCRβ+CD4+IL-17A+cells.
Western blot showing the degree of phosphorylation of Flag-Ago3 WT and the 8SA mutant in Siwi-depleted cells (Siwi KD).
F. Sister chromatid exchanges in cells that were not treated (NT) or exposed to 2 μM olaparib (OLA) for 16 hrs. Black arrows in images show SCEs in olaparib-treated cells. Scale bar: 5 μm. N=2.
(B+C) Positive charged amino acid residues at the C and negative charges at the N terminus make EGFL7 a polar molecule with a preference towards oligomerization
(A) U2OS cells (WT) and U2OS H2AX knockout cells reconstituted with empty vector (H2AX KO+EV), wild type H2AX (H2AX KO+WT), or the E141A mutant H2AX (E141A) were treated with H2O2 or MMS. Cell viability was measured after 24 hours using MTT assay. Values are mean ± SD of three assays.
A. Immunofluorescence of SATB2 (green) and the GSC marker SOX2 or OLIG2 (red) on frozen tissue sections of human GBM surgical specimens. SATB2 is preferentially expressed by GSCs in human GBMs. Scale bar, 25 μm. B. Quantification of the fraction of SATB2+ cells in SOX2+ or OLIG2+ cells in human GBMs. More than 90% SOX2+ or OLIG2+ cells showed SATB2 staining. n=3 GBMs.
J, K Panc‐1 stable cells with GOT2 knockdown and re‐expressing the indicated proteins were seeded in a 6‐well plate, and the cells were maintained under the condition of low glucose (0.5 mM, J) or high glucose (12 mM, K). Culture medium was refreshed every day, and cell numbers were counted every 1-2 days over a period of 7 days.
(H,I) Dnmt1 immunohistochemistry in spinal cord sections from AR-97Q mice treated with or without RG108 (n = 3 per group).
(C) Representative images and PIP quantification. The number of samples per group for each experiment are included within each bar graph.
HEK293T cells transfected with TREM2 or AD variant incubated with 500 nM Aβ42 monomers were lysed for Aβ42 quantifications by ELISA. Expression of TREM2 increases Aβ42 level detected compared to mock control as well the AD variant. Results were averaged ± standard error (n = 9, # p < 0.05 compared with TREM2, ## p< 0.01 compared with TREM2, *** p< 0.001 compared with mock, **** p <0.0001 compared with mock, ANOVA, Tukey's Multiple Comparison Test, see Appendix table S3 for p value)
(A, B) The relative spindle angle between the polarity axis (perpendicular to the apical surfaces stained wit Bazooka (B)) and spindle axis (defined by γ-tubulin-stained centrosomes) was determined in wild type (w1118) and alix NBs (B) incubated at room temperature (RT) or ice for 40 minutes. The graphs show the percentage of FECs or NBs displaying relative spindle angle 90-75° versus 75-0° or 0-15° versus 15-90°, respectively. The graphs represent the results from three separate experiments analyzin 32 w1118-NBs-RT (dark blue bars), 64 w1118 NBs-ice (light blue bars), 56 alix1 NBs-RT (dark grey bars) and 63 alix1 NBs-ice (light grey bars) (B
A. Left panel, part of the BH3-in-groove dimer structure is shown with Gly108 from one monomer presented in sphere form, and Gly67 and Leu70 from the other monomer that have van der Waals contacts with the Gly108 presented in stick form. Asp68 and Asp71 from the other monomer are also presented in stick form. Right panel, the same structure is shown with the Gly108 changed to glutamate (G108E), which results in steric clashes with the Gly67, Leu70 and Asp71, and electrostatic repulsions with the Asp68 and Asp71.
F. Quantitative immunostainings analysis indicates that proliferation and apoptosis markers are affected by anti-miR-10b treatment. No significant effect on invasion of intracranial GBM8 was observed.
Histology: all metastatic lesions are composed of poorly differentiated cells with an epithelial 'flavour'. The neoplastic population is mainly arranged in solid nests and sheets with focal rudimental gland formation. Scale bar: 50µm.
with digitonin (mock) or digitonin and a lysate from indicated S. flexneri strains (pHldA/C means plasmid-encoded HldA and HldC) D) IL-8 production at 6 h in cells were treate
(G) Quantitative RT-PCR gene expression analysis of BCL11B in control (black bar), GATA3 shRNA or TCF7 shRNA (grey bars) transduced CD34+ thymocytes after 48h of coculture on OP9-DLL1 stromal cells, relative to the geometric mean of ACTIN and GAPDH and relative to the control-transduced sample (n=6 independent experiments with different donors for GATA3 shRNA, n=4 independent experiments with different donors for TCF7 shRNA, error bars indicate SEM and * indicates p<0.05 in a paired t-test).
(I) Volcano plot of NEF-induced changes to BHEL cell transcription. NEF-expressing or control CD4OT-II cells were co-cultured with BHEL cells in the presence of HEL-OVA for 4h before isolation of BHEL cells and transcriptome analysis by microarray. Shown are mRNAs with log2fold change against -log10 p-value. Out of around 20,000 genes investigated, 564 genes were induced and 977 genes were suppressed by NEF (see Dataset EV1), with 187 genes induced and 407 suppressed by NEF in a statistically significant manner.
C Representative micrographs of H&E stained histological sections of MM134 xenografts from 3 different female mice 5 months after intraductal injection. Arrows point to indian files. Scale bars, 100 μm.
D Integrated analysis of six different naïve-primed datasets. Circular and triangular points in the PCA plot represent naïve and primed samples, respectively. Each color indicates one dataset. Black points represent data generated in this study. Each dataset is indicated by the name of the corresponding author and the year of publication (Dataset EV2).
(c) The volcano plot depicts the fold changes (x-axis) and the -log10 p-values (y-axis) derived from the differential gene expression analysis using diffxpy between ILD and controls within the subset of PDGFRB+ pericytes (outlier values truncated).
(A) IF of CBX2 and SUMO2/3 in PN0 (n=10) and PN5 (n=12) zygotes. Data information: Scale bars, 10µm.
H Immunofluorescence staining against NANOG shows lack of expression in E10.5 human-mouse chimera tissue sections. Scale bars: 200 and 100 µm in the inset.
While an increased number of cytosolic vesicles can be observed at 1-SKO AZs, reduced numbers of small vesicles are present in a 1 µm radius around the ribbon of 1/2-DKOs. RA-SVs were excluded from this quantification.
(F, G) Hes5 immunoreactivity of spinal motor neurons in AR-97Q mice treated with or without RG108 (n = 3).
C. qPCR analysis of SATB2 mRNA expression in GSCs and matched non-stem tumor cells (NSTCs) (n=5).
(A) Representative hematoxylin and eosin (HE) staining of ileum. Paneth cells with typical eosinophilic granules (black arrows) on HE-stained sections at the base of crypts in Villin-Cre and COPS8-lox alleles expressing (COPS8fl/fl ), but not COPS8 knockout (KO) (COPS8ΔIEC) epithelium. Scale bar 50 μm, n=7 biological replicate per genotype.
Figure 1: Impaired fear extinction precedes memory decline in Fmn2 mutant mice.B. Fear extinction in 3-month old APPPS1 mice is similar to a non-transgenic control group (n=9/group).
(A, Gene Set Enrichment Analysis (GSEA) identifies Stem-cell like expression signature in KL tumors compared with KrasG12D/+ (K) lung tumor (A) Significance was calculated by permutation test.
e, Kaplan-Meier analysis comparing PDAC-free survival of Atg5+/+ (blue) or Atg5−/− (green) KrasG12D/+ p53−/− mice.
c and d. Representation and quantification of cell length in epidermal (e) and cortical (c) cell files. Optical, longitudinal sections of 5 DAG old Col-0 roots 12 HAT to ammonium (C) or nitrate (D) supplemented media. The first 20 epidermal (e) and cortex (c) cells (from quiescent center (QC)) are highlighted in grey and in red on ammonium (C), and in blue and green on nitrate (D), respectively. Scale bar=30 µm. Column bars denote the geometric mean of the cell lengths at the respective positions. Lines represent a polynomial regression fit, with calculated slopes between cells 10 and 20 of 3.32639+0.17172 (ammonium, epidermis), 1.22033+0.08754 (ammonium, cortex) and 1.70502+0.09532 (nitrate, epidermis), 0.82342+0.06973 (nitrate, cortex). Data are derived from 3 biological replicates, n=18 roots in each case.
A- CTD shortening leads to a decreased Pol II occupancy in promoter-proximal region. Median coverage plot of PRO-seq signal at genes encoding major isoforms. Solid lines represent median signal and shaded area refer to 95% bootstrap confidence intervals. Two independent biological replicates were analyzed.
(d) Relative acetylation (normalized to day 1) of histone H3 Lys 18 residue of chronologically ageing wild-type cells. Data represent means ± s.e.m. (n = 3).
(g) The protein features in BALF were used to train a random forest algorithm to predict lung function. The model was tested on transcriptional signatures from single cell RNA-seq data to correctly predict reduced lung function in end stage lung fibrosis when compared to controls.
(F) Bar graph shows the percentage of living (TUNEL-negative) and dyeing (TUNEL-positive) cells under various conditions tested. Higher percentage of TUNEL-positive cells is observed in CCB02-treated cultures. Data represent mean ± SEM of three independent experiments. (N)=3, (n=500 cells per condition). p values were obtained using Ordinary One-way ANOVA test *P < 0.01.
MTT assay of vector control or INSL5 overexpressing CNE1, CNE2 and HK1 NPC cell lines (upper panel) either transfected with control siRNA (NC) or GPCR142 siRNA (#1 and #2) (lower panel). n=4 biological replicates for CNE1 and CNE2 cell line, n=6 biological replicates for HK1.
C Far-UV CD spectra at 20°C (top) and thermal denaturation monitored at 222 nm (bottom) for Myc (red) and Max (black) bHLH-LZ constructs (at 16 μM and 8μM, respectively), alone or in combination (Myc+Max), as indicated. SUM Myc+Max: theoretical sum of the individual Myc and Max curves. The Myc variant used (WT, RA or HEA) is indicated at the top. The increased ellipticity signals at 222 nm and increased melting temperatures of the experimental Myc+Max mixtures (blue) compared to the theoretical sums (grey) demonstrate that MycWT, MycHEA and MycRA heterodimerize with Max to comparable extents.
An aligned view as in D, showing S1 site interactions between human OTUB1 (green) and Ub (red) (PDB 4DDG). Ub is rotated 21˚ relative to the EschOTU-Ub structure, but maintains similar primary and secondary contacts.
D. Immunostaining of endogenous mTOR and LAMP1 in C2C12-Vector and Aster-C KO cells under nutrient starvation (-AA) and in response to AA stimulation (+AA). LAMP1 was immunostained as a lysosome marker. Arrows highlight the co-localization of mTOR with LAMP1. Scale bar, 40 μm (Row 1) and 10 μm (Rows 2, 3, 4).
(I) Mitochondria from the indicated yeast strains (pex19Δ, pex19Δmsp1Δ, and pex13Δmsp1Δpex19Δ) expressing Pot1-V5 (under its endogenous promotor) were either left untreated (nt) (lane 1,4,7) or were treated with proteinase K in high fidelity buffer (PK-HF) (cutting control, lane 2,5,8) or low fidelity buffer (PK-LF) (protease protection assay, lane 3,6,9) and separated by SDS-PAGE and immunoblotted for Mia40 (intermembrane space) (α-Mia40), Pot1-V5 (α-V5), porin (outer membrane) (α-porin) and Tom22 (outer membrane but partially proteinase exposed) (α-Tom22).
(A) Volcano plot (log10) of the most enriched/decreased proteins in the mitochondrial proteome of pex3Δ versus pex3Δmsp1Δ strains as detected by quantitative mass spectrometry (n=3, biological replicates, n=1, technical replicate). 2010). Effect sizes were calculated for Cohen's d A protein's relevance to the model was determined using a p-value threshold of < 0.05 with an absolute effect size threshold of 1 or greater (large effect). Gray dots represent all measured proteins. Red dots highlight peroxisomal-associated proteins.
Representative Western blot of total (mCherry and untagged), cytoplasmic TDP-43 and nuclear levels with and without HDAC6 inhibitor.
F. Kaplan-Meier survival curves of mice intracranially implanted with T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n=8 mice; shNT+FOXM1: n=7 mice; shSATB2: n=6 mice; shSATB2+ FOXM1: n=8 mice). Median survival: shNT, 30 days; shNT+FOXM1, 28 days; shSATB2, 50.5 Days; shSATB2+FOXM1, 34 days. Ectopic expression of FOXM1 in GSCs expressing shSATB2 markedly attenuated the increased survival of mice bearing the GSC-derived GBMs.
(h) HeLa cells stably interfered for AMBRA1 were transfected with FLAG-TRAF6 and Ub-Lys-63-HA as indicated. Protein extracts were immunoprecipitated using anti-ULK1 antibody.
B Representative confocal images of the vHIP stained with mCherry (red) and c-Fos (green). C Quantification of mCherry and c-Fos double positive cells out of total mCherry-positive cells in the vHIP CA1 regions. Scale bar, 50 μm. Mock (n= 5 mice) vs. ZIKV (n= 5 mice), p=0.0013. Data information: Data were presented as mean ± SEM. *P<0.05, **P<0.01, Two-tailed unpaired t-test unless described otherwise.
(B-D) Surface biotinylation assays verified increased surface expression of αV and β3 upon appli­cation of E7 but not Fn (n = 3; one-way ANOVA)
(J) Hes5 mRNA levels in the spinal cords of wild-type, AR-24Q, and AR-97Q mice (n = 3).
Calu6 (mesenchymal) and H1975 (epithelial) NSCLC cell lines after Plk1 and INTB1 silencing with siRNA for 48 hours. Cells were then harvested, and lysates were immunoblotted for the indicated proteins.
(A) IFIT-1 reporter activity from THP-1-IFIT-1 cells transduced for 24 h with HIV-1 GFP containing 0 % (WT) or 75 % ΔCA-SP1 mutant (1 U RT/ml) in the presence of DMSO vehicle, 5 μM neviripine or 10 μM raltegravir.
C. MiR-10b inhibition induces cleavage of caspases 3 and 7 in GSC, as determined by Western blot analysis at day 5 after transfection with the inhibitor. The signals were quantified using ImageJ and normalized to beta-actin.
F Representative images of recombined (GFP+) neurons co-expressing Prox1 (grey) as a marker for neuronal fate and the stage specific markers DCX (red) for immature neurons and Calbindin (red) for mature neurons. Arrows and arrowheads indicate marker negative and marker positive cells, respectively. Scale bar = 10µm. G Fraction of DCX expressing neurons is slightly reduced in β-catex3 iDCX at 3dpT [p=0.0338], while fraction of Calbindin expressing cells is slightly increased in β-catex3 iDCX at 3dpT [p=0.0345]. At 13dpT no difference is visible in stage specific marker expression [DCX p=0.8564, Calbindin p=0.0841] (3dpT: control: n= 12 animals, β-catex3 iDCX: n= 12 animals, 13dpT: control: n= 10 animals, β-catex3 iDCX: n= 14 animals).
(o) Notch1 expression (MFI ± SEM) in HE-derived CD43+CD45+ cells at day 3 of subculture with or without DCA (3 mM) was assessed by flow cytometry (n=2 biological replicates with 1-2 technical replicates, unpaired t test). Data information: ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001
(D) Representative images showing effect of MBELNs with and without TCDD (1 nm)/ carbobenzoxy-Leu-Leu-leucinal (MG132; 20 µM) on co-expression of AhR and COPS8 using immunofluorescence. Scale bar 10 μm, images are from three biological replicates.
(D) LLC tumor-bearing mice were pretreated with IgG or AB-Tie1-39. On day 14, lung tissue lysates were immunoblotted with anti-phospho-TIE2, total-TIE2 and ACTB (upper panel; n = 6 mice). Densitometric quantitation of the blots presented in the upper panel is shown (mean ± SD, n = 6 mice). **, P<0.01 (two-tailed Mann-Whitney U test).
A Experimental scheme to analyze the impact of β-catenin signaling activity on adult-born neuron development in middle-aged mice. Tamoxifen (TAM) was applied i.p. every 12 hours for five days. Animals were analyzed 3 and 13 days post Tamoxifen (dpT).
(a) Trafficking of neutrophils (N), classical monocytes (cMOs), non-classical monocytes (ncMOs), B lymphocytes, CD4+ T lymphocytes, and CD8+ T lymphocytes to the peritoneal cavity as assessed 6 h after intraperitoneal stimulation with recombinant murine uPA, tPA, PAI-1, uPA-PAI-1, or tPA-PAI-1 in WT mice by multi-channel flow cytometry. The gating strategy and quantitative data are shown (mean±SEM for n=5 mice per group; #p<0.05 vs. saline; One-way ANOVA).
Model of the regulation of ferritin fate by NCOA4 in cellular iron homeostasis. Under iron depletion, ferritin is degraded by the NCOA4 dependent macroautophagy pathway. In the early phase of iron treatment, NCOA4 forms condensates that sequester ferritin for ferritin accumulation. During prolonged iron treatment, NCOA4 condensates bind ferritin, forming a complex that is trafficked to the lysosome in a TAX1BP1-dependent manner to avoid iron deficiency.