EMBO/sd-character-level-ner huggingface dataset:
Figure 5A. Exemplary normalized current responses a -120mV pulse showing fast Ca2+ dependent inactivation (FCDI). B. Percentage FCDI (plateau/peak, mean±s.e.m.), for HEKS1/S2-/- cells transfected with STIM1+ORAI1 (O1 S1), 22 patch-clamped cells or transfected with STIM1A and ORAI1 (O1 S1A,17 cells) from three independent transfections. Data information: , *P<0.05, **P<0.01, unpaired T test with Welch's correction. For patch-clamp experiments, number in bars indicate measured cells, which were from three independent transfections. C. Docking model of the STIM1 CAD domain to ORAI at the beginning (red) and at the end (green) of a 50ns MD simulation. D. close up of interacting residues. E. alternate docking approach. F. Traces showing average changes (mean±s.e.m.) in intracellular Ca2+ (Fura2 ratio) over time in response to perfusion of different [Ca2+]o as indicated in the upper bar with constructs as indicated expressed in HEKS1/S2-/-. G. Quantification of the relative reduction of ratio/time or ratio for cells (135 H Data information: , ***P<0.001, Kruskal-Wallis Anova with Dunn's multiple comparisons test Data points (total n) were obtained from three biological replicates with each three technical replicates (with multiple cells) and shown as mean±s.e.m for traces (F) and as scatter plots with the underlying boxes showing the means for individual parameters (G). H,I. Average traces (mean±s.e.m.) showing whole- cell current density (CD) over time extracted at -80 mV in in HEKS1/S2-/- cells co-transfected either with STIM1 (black) or STIM1A (red) and ORAI1 (H) or with ORAI1 R77E (I) and recorded using extracellular solution containing 2 mM Ca2+. J. Average maximum CDs recorded from cells measured in [H,I] (n within bars). Data information: I , *P<0.05, **P<0.01, unpaired T test with Welch's correction. For patch-clamp experiments, number in bars indicate measured cells, which were from three independent transfections.
Figure 3A-D. mRNA expression of inflammation markers relative to LysM-Cre/Nrp1+/+ in mouse RPE-choroid-sclera complexes at D3 for Il1b (A); n = 10 (LysM-Cre/Nrp1+/+ and LysM-Cre/Nrp1fl/fl), Il6 (B); n = 7 (LysM-Cre/Nrp1+/+ and LysM-Cre/Nrp1fl/fl), Vegfa (C); n = 10 (LysM-Cre/Nrp1+/+ and LysM-Cre/Nrp1fl/fl), Tnf (D); n = 5 (LysM-Cre/Nrp1+/+), n = 10 (LysM-Cre/Nrp1fl/fl). mRNA expression of inflammation markers relative to LysM-Cre/Nrp1+/+ in mouse RPE-choroid-sclera complexes at D7 for Il1b (E); n = 6 (LysM-Cre/Nrp1+/+), n = 10 (LysM-Cre/Nrp1fl/fl), Il6 (F); n = 3 (LysM-Cre/Nrp1+/+), n = 4 (LysM-Cre/Nrp1fl/fl), Vegfa mRNA expression of inflammation markers relative to LysM-Cre/Nrp1+/+ in mouse RPE-choroid-sclera complexes at D7 for Il1b (G); n = 6 (LysM-Cre/Nrp1+/+), n = 10 (LysM-Cre/Nrp1fl/fl), Tnf (H); n = 8 (LysM-Cre/Nrp1+/+), n = 12 (LysM-Cre/Nrp1fl/fl). I. Representative Western Blot showing pNF-κB expression in LysM-Cre/Nrp1+/+ (Ctrl) and LysM-Cre/Nrp1fl/fl (Nrp1fl/fl). J. Quantification of pNF-κB expression in LysM-Cre/Nrp1+/+ and LysM-Cre/Nrp1fl/fl BMDM; n= 6. K, L. mRNA expression relative to LysM-Cre/Nrp1+/+ of inflammation markers in mouse BMDMs for Il1b (K); n = 4 (LysM-Cre/Nrp1+/+), n = 5 (LysM-Cre/Nrp1fl/fl) and Il6 (L); n = 3 (LysM-Cre/Nrp1+/+), n = 2 (LysM-Cre/Nrp1fl/fl) M. Representative FACS plots of M1 and M2-Like macrophages in LysM-Cre/Nrp1+/+ and LysM-Cre/Nrp1fl/fl BMDMs. N, O. Quantification of M1-Like macrophages (F4/80+, CD11b+, CD11c+, CD206-)(N), M2-Like macrophages (F4/80+, CD11b+, CD11c-, CD206+) (O) in LysM-Cre/Nrp1+/+ and LysM-Cre/Nrp1fl/fl BMDMs relative to LysM-Cre/Nrp1+/+; n= 5. P. Heatmap (left) and enrichment plot (right) of GO Angiogenesis gene set enrichment analysis (GSEA) of wildtype and LysM-Cre/Nrp1fl/fl peritoneal macrophages; n = 2. NES, normalized enrichment score; FDR, false discovery rate.
linc-MYH keeps the number of MuSCs in check and limits muscle fiber growth A, B Body weight (A) of male linc-MYH KO and ctrl mice at 10 weeks (n = 24 KO/17 WT; Student's t test, two-tailed, **p < 0.Figure 2Body weight (A) of male linc-MYH KO and ctrl mice at 10 weeks (n = 24 KO/17 WT; Student's t test, two-tailed, **p < 0.01) tibia length (B) of male linc-MYH KO and ctrl mice of 10 weeks (n = 9 KO/13 WT; Student's t test, two-tailed, not significant). Data are mean ± S.E.M. C, D Weight of m. tibialis anterior (TA) and m. digitorum longus (EDL) of male linc-MYH KO and ctrl mice at 10 weeks (n = 9 KO/10 WT for TA, n = 10 KO/11 WT for EDL; Student's t test, two-tailed, **p < 0.01, ***p<0.001). Data are mean ± S.E.M. Size distribution of fibers in cross sections of TA (E, F) and EDL (G, H) muscle in male linc-MYH KO (blue) and ctrl (grey) animals at 10 weeks. A significant increase of fiber size was observed for TA and EDL muscles of linc-MYH KO mice (n = 5 KO/4 WT for TA, n = 3 KO/3 WT for EDL muscle, two-way ANOVA with Fisher's multiple comparisons; *p < 0.05, **p < 0.01; > 250 fibers per animal were counted). Data are mean ± S.E.M. Size distribution of fibers in cross sections of TA and EDL muscle in male linc-MYH KO (blue) and ctrl (grey) animals at 10 weeks. A significant increase of fiber size was observed for TA and EDL muscles of linc-MYH KO mice (n = 5 KO/4 WT for TA, n = 3 KO/3 WT for EDL muscle, two-way ANOVA with Fisher's multiple comparisons; *p < 0.05, **p < 0.01; > 250 fibers per animal were counted). Data are mean ± S.E.M. K Numbers of myonuclei in cross sections of TA and EDL muscle (TA: n = 6 KO/6 WT, EDL n = 3 KO/3 WT animals, Mann-Whitney test one-tailed, **p<0.01, *p = 0.05) Data are mean ± S.E.M. L-O Number of myonuclei / fiber and of Pax7pos (red) MuSCs (red arrows) on myofibers isolated from flexor digitorum brevis muscle. Nuclei were stained using DAPI (blue). Scale bar in L indicates 50 µm for L and M. (N) The number of myonuclei / isolated fiber is increased in linc-MYH KO compared to WT myofibers (n = 4 KO/4 WT, > 26 fibers/animal; Mann-Whitney test two-tailed, * p < 0.05). (O) Number of MuSCs on isolated fibers from linc-MYH KO and WT mice (n = 4 KO/4 WT animals, Mann-Whitney test two-tailed; ****p < 0.0001). Data are mean ± S.E.M. P-S Number of Pax7 (red) positive MuSCs in cross sections of tibialis anterior muscle (P-R); the relative amount of quiescent MuSCs identified by double staining (yellow arrows) for Pax7 and CalcR (green) does not significantly differ between WT and linc-MYH KO animals (S). Nuclei were stained with DAPI (blue). Scale bar in P indicates 25 µm for P and Q. (n = 3 KO/3 WT animals, Mann-Whitney test one-tailed; *p = 0.05, ns: not significant). All data are mean ± S.E.M. T The number of MuSC-related events in FACS experiments is significantly increased in linc-MYH-KO animals (n = 7 KO/7 WT, Student's t test, two-tailed, *p = 0.0126). Data are mean ± S.E.M. U-V Structure of MuSCs and the heterochromatin content of MuSC nuclei is not changed in linc-MYH KO (V) compared to WT mice (U).
Proposed model for a role of NlpI in coordinating formation of PG multi-enzyme complexes containing EPases.Figure 6A. Estimated number of molecules and molarity of PBP1a/LpoA and EPases. 1Numbers obtained by ribosomal profiling in rich growth medium (Li et al., 2014). 2Concentration of monomer. 3Decreases in the presence of NlpI (Singh et al., 2015). 4"no change detected". The periplasmic concentrations of proteins were estimated for a cell with periplasmic volume of 3.33 ×10-16 L, where 1 molecule corresponds to 5 nM. B. Hypothetical model of NlpI scaffolding endopeptidases during cell elongation. Black arrows indicate interacting proteins with numbers indicating apparent EC50 / KD values. OM, outer membrane; CM, cytoplasmic membrane. MepS-PBP1A interaction is not represented due to illustrative restrictions.
Figure.Figure 5(A) RNA-seq was performed with RNAs extracted from ASCs (FACS purified and cultured for 36hrs) of YY1iKO or Ctrl mice; scatter plot shows differentially expressed genes with a fold change >=2 (red dots) in YY1iKO vs Ctrl. (B) Gene ontology (GO) analyses of the above up-regulated genes. The top 10 enriched GO terms are displayed on the Y axis and adjusted P values on the X axis. (C) Genomic snapshots showing the examples of mitochondrial genes (Uqcr11, Mrp11, Apo5o and Nme4) up-regulated in YY1iKO vs Ctrl cells. (D) GSEA analysis shows oxidative phosphorylation gene set is enriched in YY1iKO vs Ctrl cells (p-value < 0.0001). (E) RT-qPCR validation of the expression of the selected up-regulated mitochondrial genes in YY1iKO vs Ctrl. (F) Protein levels of the respiratory chain complexes I-V components in YY1iKO vs Ctrl ASCs were detected using a mitoprofile antibody cocktail. β-actin was used as the loading control. (G) RNA-seq was performed using RNAs from freshly isolated SCs (FISCs); scatter plot shows differentially expressed genes with a fold change >=2 (red dots) in YY1iKO vs Ctrl. (H) GO analysis for the up-regulated genes in YY1iKO vs Ctrl. (I) Genomic snapshots show the selected mitochondrial genes (Cox8b, Nme4, Slc25a29, Nnt) up-regulated in YY1iKO vs Ctrl FISCs. (J) ChIPseq was performed using chromatins from WT ASCs and the genomic distribution of 2031 YY1 binding peaks is shown. (K) De novo motif prediction by DREME revealed the enrichment of canonical YY1 motifs in the above binding regions. (L) Venn diagrams show the overlapping (90 genes) between the above identified binding target genes (1387) and the upregulated genes from ASC RNAseq (979). (M) GO analysis of the above 90 genes revealed an extreme enrichment of mitochondrial related terms. (N) Genomic snapshots of two of the above identified mitochondrial genes (Tomm40I and Ndufa13) that are bound by YY1 in their TSS (ChIPseq tracks) and up-regulated by YY1 deletion (RNAseq tracks). (O) ChIP-qPCR validation of YY1 binding on some mitochondrial genes. Negative control (NC) represents a genomic region on Chromosome 11 with no YY1 binding peak identified. Enrichment fold was calculated as the amount of amplified DNA from YY1 binding sites normalized to values obtained from IgG control. ChIP-qPCR was performed to show the enrichment of Ezh2 (P) binding on selected mitochondrial genes. ChIP-qPCR was performed to show the enrichment of Suz12 (Q) binding on selected mitochondrial genes. ChIP-qPCR was performed to show the enrichment of H3K27 me3 (R) binding on selected mitochondrial genes. (S) A construct to express wild type YY1(WT), or two mutants lacking DNA binding activity, YY1(1-394) and YY1(S365D) were transfected into YY1iKO cells; the expression of mitochondrial gene was measured. Error bars represent s.d.'s of the mean. Student's t-test (two-sided): N.S = non-significant, *P<0.05, **P<0.01, ***P<0.001.
Rab11-FIP3 regulates T cell spreading via a Rac1-dependent mechanism.Figure 6Jurkat T cells were transfected with control or FIP3.1 siRNA, then transfected again with empty vector or with expression vectors encoding Myc-tagged-Rac1WT, -Rac1G12V (constitutively active mutant), or -Rac1Rac1T17N (dominant negative mutant). Cells were allowed to spread on poly-lysine-coated coverslips for the indicated time and the spreading area of cells measured. A-C, H: Population analyses of the spreading area of cells at the level of the contact surface. A-C: Time course plots representing the mean ± SEM of the data shown as in I. D-G: Staining for nucleus (blue = Dapi), F-actin (green = phalloidin) and Rac1-Myc (red = anti-Myc) is shown. A single optical section at the level of the contact surface is shown. Bar, 5 µm. H: Dot plots corresponding to measurements at 15 min of spreading: Each dot represents one cell. Horizontal bars represent the mean ± SEM. Mann-Whitney test. Data are representative of three experiments.
Figure 2Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation. (A) Upper part, scheme of S. pombe sexual differentiation. Lower part, microscopy images showing WT and mmi1∆ cells, and mmi1∆ cells expressing Mmi1-R351E or Mmi1-R381E mutant proteins. Images were taken after 24 hours of induction of sexual differentiation by growth on SPAS medium. The percentage of cells that underwent differentiation and iodine vapor assays, conducted on the corresponding patches, are shown at the bottom of each image. Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation. (B) Mating assay showing the percentage of zygotes forming over time in the same cells as in (A). Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation.(C) Scheme of nam1-byr2 locus. Mmi1 UNAAAC binding motifs are depicted by white lines. (D) Scheme of nam1-byr2 locus in nam1-1 cells highlighting the 8 UNAAAC motifs mutated (red lines). Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation. (E) Mmi1 RNA-IPs showing the specific loss of binding of Mmi1 to nam1-1 lncRNA but not to mei4 mRNA, another target of Mmi1. Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation. (F) Mating assay showing the percentage of zygotes formed over time in WT and nam1-1 cells. Average fold enrichment is shown with error bars that indicate mean average deviations (n=3) (B, E and F). Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation. (G) Western blots showing the level of Flag-Byr2 protein over the first 4 hours of sexual differentiation in WT and nam1-1 cells. Tubulin (Tub1) level was used as a loading control. Figure 2. Mmi1 binding to nam1 lncRNA controls MAPK-mediated entry into sexual differentiation. (H) Microscopy images of WT (h90) cells transformed with an empty plasmid (Control) and nam1-1 cells transformed with either an empty plasmid (Control) or a plasmid expressing Byr2 protein (Byr2), after 24 hours of induction of sexual differentiation. Scale bar, 10 µm.
Cycline signature analysis in METABRIC dataset.Figure 2A. Primary human breast cancers of the METABRIC dataset were stratified according to high or low YAP activity signature [47] and by TP53 mutational status, and then the levels of the cycline signature score were determined in the four groups. Cyclin activity is significantly higher in mut-p53 tumors with high levels of the YAP signature, as visualized by box-plot. Signature scores have been obtained summarizing the standardized expression levels of signature genes into a combined score with zero mean [7]. The values shown in graphs are thus adimensional. The bottom and top of the box are the first and third quartiles, and the band inside the box is the median; whiskers represent 1st and 99th percentiles; values lower and greater are shown as circles (p<0.0001, n=701). B. Same as in A) using the YAP/TAZ activity signature of Zhang et al. (2009) (p<0.0001, n=701). C. Kaplan-Meier analysis representing the probability of disease-specific survival in mutant and wild-type p53 breast cancer patients from the METABRIC dataset stratified according high or low YAP/TAZ signature score. The log-rank test p-value reflects the significance of the association between high levels of the YAP/TAZ signature score and shorter survival in in mutant-p53 as compared to wild-type p53 patients (p<0.0001, n=251). All p-values were calculated with two-tailed Student's t-test.
Figure 1SARS-CoV-2 complete genome sequence was used to design eight overlapping subgenomic viral fragments covering the complete genome. Positions on the genome (in nucleotide) are indicated in bold red.
Identification of a Nrd1-Interaction Motif (NIM) in Sen1 that is critical for the integrity of the NNS-complex.Figure 1A) Deletion of the CID domain dramatically reduces the interaction of Nrd1 with Sen1. Coimmunoprecipitation (CoIP) experiments using TAP-tagged Nrd1 (either wt or ∆CID) as the bait. Representative gel of one out of two independent experiments. B) Scheme of Sen1 protein. Globular domains are denoted by solid bars whereas intrinsically disordered regions are shown by a line. The disorder prediction was obtained using IUPred (Dosztányi et al., 2005). The sequence of the RNAPII S5P-CTD and Trf4 and Sen1 NIMs is shown on the top. Structural elements that are important for the interaction with Nrd1 CID are indicated. Conserved positions are underlined. C) Deletion of the NIM decreases substantially the association of Nrd1 with Sen1. CoIP experiments using Nrd1-TAP as the bait in a SEN1 or sen1∆NIM background. Representative gel of one out of two independent experiments. D) CoIP experiments using HA-tagged Sen1, either wt or ∆NIM, as the bait. Representative gel of one out of two independent experiments. Protein extracts were treated with RNaseA prior to immunoprecipitation. In these experiment, Sen1 could not be detected in the input extracts.
Figure 6(A) RT-qPCR and Western Blot analyses of CaSR expressions in MC3T3-E1 cells treated with IL-1β. n=3, one‐way ANOVA, Bonferroni post hoc. (B) RT-qPCR and Western Blot analyss of CaSR expressions in MC3T3-E1 cells treated with TNFα. n=3, one‐way ANOVA, Bonferroni post hoc. (C) RT-qPCR and Western Blot analyses of CaSR expressions in MC3T3-E1 cells treated with IL-17A. n=3, one‐way ANOVA, Bonferroni post hoc. (D) RT-qPCR and Western Blot analyses of CaSR expressions in MC3T3-E1 cells treated with IL-22. n=3, one‐way ANOVA, Bonferroni post hoc. (E) RT-qPCR and Western Blot analyses of CaSR expressions in MC3T3-E1 cells treated with IL-23. n=3, one‐way ANOVA, Bonferroni post hoc.
Simulation of primary response gene dynamics upon different signalling durations.Figure 2 Figure 2 - Simulation of primary response gene dynamics upon different signalling durations. A: Different activation patterns of signalling molecules (input functions, left) can elicit multiple different response profiles (right) with different response times (r) depending on mRNA half-lives (t1/2) and transcriptional delays (∆t). Rapid induction requires short half-lives (red lines). Late induction can be caused by transcriptional delays (blue lines), long half-lives (yellow lines), or combinations thereof. Decoding of signal duration depends on mRNA half-life. Short-lived mRNAs relay signal duration to response duration, whereas long-lived mRNAs decode signal duration to response amplitude (yellow lines). B: Response amplitude for all simulated combinations of mRNA half-life and transcriptional delay. Response amplitude is shown over time (columns) and in respect to input function (rows). For sustained signalling, all primary response genes exceed their half maximum response amplitude. Pulse and transient signalling inputs are only sufficient for immediate-early genes and short-lived delayed early genes. Long-lived mRNAs with half-lives greater 120 minutes require sustained signalling inputs to exceed their half maximum response amplitude. Example parameter sets displayed in (A) are marked with asterisk in (B). Dashed lines indicate cluster borders: IEG: Immediate-early genes, t1/2 ≤ 120min and ∆t ≤ 30min. ILG: Immediate late genes, t1/2 > 120min and ∆t ≤ 30min. DEG: Delayed early genes, ∆t > 30min.
Figure 4Immunohistochemical images of the IPN region from control and TMEM16A cKO mice. The c-Fos signal was greatly reduced in TMEM16A cKO mice. Scale bar = 200 μm. A summary bar graph showing reduced c-Fos expression in the IPN in TMEM16A cKO mice. (NCTL=7 (n=42), NcKO=6 (n=36), unpaired Student t-test, data are presented as mean values ± s.e.m.). An illustration of a brain slice (sagittal view) to show the site of electrophysiological recordings. Representative traces of sEPSCs of neurons in the medial part of the IPN showing a reduced frequency in TMEM16A cKO mice compared to that of littermate controls. Summary bar graphs of the frequency and amplitude of the sEPSCs of IPN neurons in TMEM16A cKO (cKO) mice and littermate controls (CTL). Data are presented as mean values ± s.e.m. (paired Student t-test). The numbers inside each bar indicate the numbers of recorded cells.
Generation of a zebrafish chd7 mutant using CRISPR/Cas9.Figure 1A. Chromatograms showing the confirmation a 1-nucleotide insertion mutation by Sanger sequencing. B. Gross morphological analyses of control (chd7+/+; top left image), heterozygous (chd7+/-; bottom left image) and knockout mutants (chd7-/-; images in right panel). C. Kaplan-Meier Survival plot showing low survival of chd7 mutants after 12 dpf (N=5). D. Measurement of head size of control (n=12) and mutants (n=14) showing significantly smaller head size in chd7-/- fish (****p<0.0001, Student's t-test). E. Acetylated tubulin staining in 28 hpf controls (left) and mutants (right) showing severely affected outbranching of the trigeminal nerve (Vth cranial nerve). Notably, chd7-/- display reduced branching of the Vth cranial nerve (arrows) and axonal arborisation in the tectal area. Graphs showing quantitative analyses of percentage (n=5) and mean total length of peripheral projections (n=6) per zebrafish in controls and mutants (***p<0.001; **p<0.005, Student's t-test). F. Locomotor activity of control (black), heterozygous (blue) and mutants (red) showing significant hyperactivity of mutants in dark and light cycles (N=3, n=48). G. Average activity per second during dark cycle (left) is significantly increased in chd7-/- mutants compared to chd7+/+ (n=32; ****p<0.0001, Student's t-test). Representative swimming tracks during dark cycle of control and mutant fish (right). Mutant chd7-/- larvae displayed hyperactive swimming.
Expression pattern of Igsf10 mRNA in mouse and human developing brain .Figure 4A-J: Igsf10 expression was not observed at E10.5 (A, sagittal section) but readily detectable at E12.5, in the NM (B and C, frontal sections) and E14.5, along the migratory path of GnRH neurons (D, frontal section). Igsf10 positive cells were negative for Isolectin b4 (marker of vasculature) at all developmental stages (E, at E14.5). Incubation with the sense probe resulted in no signal at all stages (E14.5 shown in F). Peripherin-positive cells olfactory axons were extending within the Igsf10-positive NM (G and H). Cxcl12 mRNA expression is shown in I (E14.5, frontal section). GnRH neurons are located in the MPOA by E17.5 (J, frontal section) in an Igsf10 negative area. K-N: In human 11pcw brains, IGSF10 expression pattern was similar to that observed in mouse, with GnRH neurons interspersed in an IGSF10-positive NM (K-N, frontal sections, see also schematic in P). IGSF10 positive cells were also detected in the olfactory bulb (OB, panel O). Sense probe resulted in no specific signal (panel N).
Figure 3Schematics representing data of all Sgs1 phosphorylation sites detected in a large scale, high coverage, phosphoproteomic dataset (Lanz et al, 2021). Bolded sites indicate those subsequently detected in the IP-MS phospho-mapping experiment in Figure 3C. TIM, Top3 Interacting Motif; HRDC, Helicase and RnaseD C-terminal domain. Immunoblot analysis of Sgs1 N-terminus (amino acids 1-647) from cells treated with increasing doses of the DNA alkylating drug MMS in the absence of RAD9 and/or MEC1. This truncated Sgs1 protein was expressed from its native promoter in this and all subsequent experiments. Quantitative mass spectrometry analysis of phosphopeptides from affinity purified Sgs1 N-terminal fragment expressed in rad9∆ or wild-type 0.02% MMS-treated cells. Error bars represent standard deviation of two or more independent peptide-spectrum matches (PSMs) corresponding to the indicated sites. Rmi1, a component of the Sgs1-Top3-Rmi1 (STR) complex, is hyper-phosphorylated on S/T-Q motifs in rad9Δ cells. Model for Mec1-dependent phosphorylation of the STR complex. For simplicity, the ssDNA-binding heterotrimer RPA and the obligatory Mec1 cofactor Ddc2 have been omitted from the diagram.