EMBO/sd-character-level-ner huggingface dataset:
Figure 6(A, B) SARS-CoV-2 binding to monocyte-derived DCs (A) or primary LCs (B) in absence or presence of UF heparin (250 IU/mL) or LMWH enoxaparin (250 IU/mL). Data information: Data show the mean values and error bars are the SEM. ordinary one-way ANOVA with Tukey's multiple-comparison test. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 (A) (n=4), (B) (n=4) DCs: Dendritic cells, LCs: Langerhans cells, (C) DCs and LCs were infected with SARS-CoV-2 pseudovirus and infection was determined after 5 days by measuring luciferase reporter activity. As positive controls Huh7.5 cells were infected. Data information: Data show the mean values and error bars are the SEM. ordinary one-way ANOVA with Tukey's multiple-comparison test. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 (n=4 measured in triplicate), DCs: Dendritic cells, LCs: Langerhans cells, RLU: relative light units (D) ACE2 cell surface expression on DCs, LCs and Huh7.5. Representative data for an experiment repeated more than three times with similar results (n=3 in duplicates). Data information: Data show the mean values and error bars are the SEM. DCs: Dendritic cells, LCs: Langerhans cells, ND: Not determined. (E) Graphical overview of the cell-to-cell viral transmission assay. (F, G) DCs (F) and LCs (G) were pre-incubated with SARS-CoV-2 pseudovirus for 4 hours at 37°C in presence or absence of UF heparin (250 IU/mL) or LMWH enoxaparin (250 IU/mL), extensively washed and co-cultured with Huh7.5 cells. Transmission by DCs or LCs to Huh7.5 cells was determined by luciferase reporter activity. Data information: Data show the mean values and error bars are the SEM. (F) ordinary one-way ANOVA with Dunnett's multiple-comparison test. *P≤0.05, **P≤0.01 (n=4 measured in triplicate), (G) ordinary one-way ANOVA with Tukey's multiple-comparison test. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 (n=3 measured in triplicate). DCs: Dendritic cells, LCs: Langerhans cells, RLU: relative light units (H, I) SARS-CoV-2 isolate (hCoV-19/Italy) was pre-treated with LMWH enoxaparin (250 IU/mL) for 30 min at 37°C. DCs (H) and LCs (I) were exposed to either the untreated or pre-treated SARS-CoV-2 isolate (100 TCID/mL) for 24h, washed thoroughly and co-cultured with Huh7.5 cells. Quantification of viral RNA was measured by quantitative real-time PCR. Data information: Data show the mean values and error bars are the SEM. (H, I) ordinary one-way ANOVA with Tukey's multiple-comparison test. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 (H) (n=3 in duplicates), (I) (n=4 in duplicates). DCs: Dendritic cells, LCs: Langerhans cells
Figure 1(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). (b) Neutrophil and cMO trafficking to the peritoneal cavity elicited by recombinant murine uPA-PAI-1 as assessed in WT mice receiving neutrophil-depleting anti-Ly-6G mABs or isotype control ABs, quantitative data are shown (mean±SEM for n=4 mice per group; #p<0.05 vs. saline; *p<0.05 vs. isotype; One-way ANOVA). (c) Intravascular endothelial cell interactions and transmigration of neutrophils (Ly-6G+ CX3CR-1-; red) and cMOs (Ly-6G- CX3CR-1low; green) to the perivascular tissue as assessed 6 h after intrascrotal stimulation with recombinant murine uPA, PAI-1, or uPA-PAI-1 in postcapillary venules of the cremaster of CX3CR-1GFP/+ mice by multi-channel in vivo microscopy. Representative still images (scale bar: 50: µm) and quantitative data are shown (mean±SEM for n=4 mice per group; #p<0.05 vs. saline; *p<0.05 vs. uPA; §p<0.05 vs. PAI-1; One-way ANOVA).
The charge at the E2HTH modulates the intrinsic activity of the E2.Figure 2A Cartoon of pulse-chase diUb synthesis assays, related to (E-H), where the donor Ub is fluorescently labeled and transferred to unlabeled acceptor UbΔGG. B Crystal structure of UBE2CHTH (yellow) aligned to UBE2SHTH (green, PDB: 1ZDN) (Lin et al, 2002; Sheng et al, 2012). C Crystal structure of UBE2D2HTH (blue) aligned to UBE2SHTH (green, PDB: 1ZDN) (Ozkan et al, 2005; Sheng et al., 2012). D Crystal structure of UBE2R2HTH (red) aligned to UBE2SHTH (green, PDB: 1ZDN) (Sheng et al., 2012; Williams et al, 2019). E, F, G, H Pulse-chase diUb synthesis assays using different E2s, UBE2S (E), UBE2C (F), UBE2D2 (G), or UBE2R2 (H). DiUb formation was monitored by fluorescent scanning (top), quantitated (bottom), and assessed for statistical significance by either an unpaired Welch's t-test (UBE2C and UBE2R2) or one-way ANOVA (UBE2S and UBE2D2) (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001, n=3 independent, technical replicates for all E2s). Error bars: SEM. Data normalized to the maximal WT diUb formation. # denotes a contaminant band. I Weblogos of the HTH motifs of UBE2S, UBE2C, UBE2D, and UBE2R. Residues colored according to their average net negative (red) or positive (blue) charge. Residues of interest on UBE2S, UBE2C, UBE2D, and UBE2R are labeled. UBE2R is extended to show conservation of D143, used in Fig 4. J Negative charges at the E2HTH weakens the diUb synthesis activity of the E2 family. Electrostatic models of the UBE2S, UBE2C, UBE2D2, and UBE2R2 HTH motifs were generated using the PBEQ solver (Jo et al, 2008). Active site cysteine is indicated as a yellow sphere. Negative to positive charge represented as a red to blue gradient.
Figure 1(A) hCAP-G is composed of 1,015 amino acids and contains 19 HEAT repeats. hCAP-H is composed of 730 amino acids and contains 5 conserved motifs (I: hSMC2 binding region, II: hCAP-D2 binding region, III: DNA binding region, IV: hCAP-G binding region, V: hSMC4 binding region). (B) Scheme of the hCAP-G-H subcomplex. Residues 1-478 of hCAP-G were fused to residues 554-900 of hCAP-G. hCAP-G (1-478, 554-900) was co-expressed in E.coli and co-purified for crystallography. (C) Cartoon diagram of the crystal structure of hCAP-G (orange) in complex with a fragment of hCAP-H (green). Unstructured, disordered regions are indicated by the dots. The 19 HEAT repeats (H1-H19) and 2 disordered loops (H12 loop and H15 loop) of hCAP-G, and 4 helices (α2, α3', α3" and α4) of hCAP-H are labeled. The N- and C termini of CAP-G and CAP-H are also indicated. A molecule of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) is shown by the orange-colored stick model. A 90-degree rotated version is shown on the right. (D) Comparison of the hCAP-G-H subcomplex with its related structures. Superimposition of the structures of hCAP-G-H (orange), S. cerevisiae YCG1-BRN1 (PDB ID: 5OQQ; blue) and S. pombe CND3-CND2 (PDB ID: 5OQR; green) is presented as a Cα-tracing model. (E) Comparison of the DNA-bound form with DNA-free forms. Superimposition of the structures of DNA-bound YCG1-BRN1 (PDB ID: 5OQN; red), hCAP-G-H (orange), YCG1-BRN1 (blue) and CND3-CND2 (green)
Fig.Figure 4A. Viability IBL-202 or IBL-301 treated cells determined by CellTiterGlo. B-C. Cell cycle distribution of LU-NB-3 cells determined by flow cytometry. Bars show mean values from two independent experiments. D. Cleaved caspase 3 (Cl Casp3) levels in LU-NB-3 cells determined by western blotting. SDHA levels were used as loading control. E. Flow cytometry analyses of Annexin V and PI stainings following treatment with 360 nM IBL-202 or 50 nM IBL-301. F. Quantification of live and dead cells from the AnnexinV/PI stainings. Dead cells=PI positive, Live cells=PI negative. Dot plots from two independent experiments and error bars represent SEM.
Figure 4A. 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. B and D. The radioactive single- or double-cysteine Bax proteins with or without the indicated mutations were activated and targeted to the mitochondria that were either untreated (mito) or pretreated with NEM (NEM-mito), then oxidized and analyzed as in Figure 1B. Protein standards, and monomers and disulfide-linked dimers of the Bax proteins are indicated as in Figure 1B. n = 2 for all mutants. C. The in vitro synthesized [35S]Met-labeled Bax E69C protein with or without the additional mutations (G108E and/or S184V) were activated by the in vitro synthesized [35S]Met-labeled tBid L105C protein and targeted to the Bax-/-/Bak-/- mitochondria that were pretreated with NEM. The resulting mitochondria were isolated and oxidized by CuPhe for 0 or 30 min. The resulting "0 min" samples (1 equivalent each) and "30 min" samples (1 equivalent each) were analyzed by non-reducing SDS-PAGE and phosphor-imaging. The remaining "30 min" samples (4 equivalent each) were immunoprecipitated (IP) by either Bax- or tBid-specific antibody, and then analyzed by non-reducing SDS-PAGE and phosphor-imaging. The identities of the four major products, indicated on the right side of the image, were based on their Mr and recognition by the respective antibody. n = 2. B and D. The radioactive single- or double-cysteine Bax proteins with or without the indicated mutations were activated and targeted to the mitochondria that were either untreated (mito) or pretreated with NEM (NEM-mito), then oxidized and analyzed as in Figure 1B. Protein standards, and monomers and disulfide-linked dimers of the Bax proteins are indicated as in Figure 1B. n = 2 for all mutants.
Figure 7A. Immunohistochemistry of GFAP was performed. GFAP intensities in the cortex and the striatum were analyzed. Values were derived from three individual mice (n=3). *** P < 0.001, ** P < 0.01, * P < 0.05, one-way ANOVA. B. Immunohistochemistry of IBA-1 and MHC Class II were performed. Arrow indicates MHC class II positive cells. Numbers of MCH class II positive cells in the cortex and the striatum were analyzed. Values were derived from three individual mice (n=3). *** P < 0.001, ** P < 0.01, one-way ANOVA.
Figure 3A Representative recordings of sEPSCs from pyramidal neurons in mPFC. B Quantitation of sEPSC frequency mPFC from pyramidal neurons in mPFC. C Quantitation of sEPSC amplitude of mPFC pyramidal neurons. Mock, n=14 neurons from 4 mice; ZIKV, n= 15 neurons from 5 mice in B and C. Data information: Data were collected from PL and IL regions of mPFC, and were presented as mean ± SEM. *p<0.05, **p<0.01. Two-tailed unpaired t-test unless described otherwise. D Representative recordings of sIPSCs from pyramidal neurons in mPFC. E Quantifications of sIPSC frequency from pyramidal neurons in mPFC. F Quantifications of sIPSC amplitude of mPFC pyramidal neurons. Mock, n=14 neurons from 5 mice; ZIKV, n= 18 neurons from 6 mice in E and F. Data information: Data were collected from PL and IL regions of mPFC, and were presented as mean ± SEM. *p<0.05, **p<0.01. Two-tailed unpaired t-test unless described otherwise. G Sample traces evoked by a depolarizing current stimulation of 400 pA. H Frequency-current (F-I) plots. mock (n = 20 neurons, 5 mice) vs. ZIKV (n = 22 neurons, 6 mice), p=0.0140. Data information: Data were collected from PL and IL regions of mPFC, and were presented as mean ± SEM. *p<0.05, **p<0.01. Two-tailed unpaired t-test unless described otherwise. I Representative traces from CPn neurons in response to a hyperpolarizing current. J Action potential frequencies of CPn neurons evoked by accelerating current injection. 200 pA: mock n = 18; ZIKV n = 21 neurons, p=0.1003; 400 pA: mock n = 18; ZIKV n = 21 neurons, p=0.0085; 600 pA: mock n = 12; ZIKV n = 18 neurons, p=0.0169. Data information: Data were collected from PL and IL regions of mPFC, and were presented as mean ± SEM. *p<0.05, **p<0.01. Two-tailed unpaired t-test unless described otherwise. K Representative traces from COM neurons in response to a hyperpolarizing current. L Action potential frequencies of COM neurons evoked by accelerating current injection. 200 pA: mock n = 20; ZIKV n = 20 neurons, p=0.2705; 400 pA: mock n = 15; ZIKV n = 15 neurons, p=0.0345; 600 pA: mock n = 12; ZIKV n = 14 neurons, p=0.0160. Mock n=7; ZIKV n=8 mice in J and L. Data information: Data were collected from PL and IL regions of mPFC, and were presented as mean ± SEM. *p<0.05, **p<0.01. Two-tailed unpaired t-test unless described otherwise.
CTD shortening in human cells alters transcribed putative enhancers.Figure 4A- Pie chart showing numbers of putative eRNAs annotated using TT-seq data in RPB1-52R and RPB1-25R cells. B- Box plots showing RNA synthesis levels (RPKs) of putative eRNAs annotated using TT-seq data in RPB1-52R and RPB1-25R cells. p value =9.27e-103 (Mann-Whitney U test) Box limits are the first and third quartiles, the band inside the box is the median. The ends of the whiskers extend the box by 1.5 times the interquartile range. Two independent biological replicates were analyzed. C- Box plots showing lengths of putative eRNAs annotated using TT-seq data in RPB1-52R and RPB1-25R cells. p value =0.000519 (Mann-Whitney U test) Box limits are the first and third quartiles, the band inside the box is the median. The ends of the whiskers extend the box by 1.5 times the interquartile range. Two independent biological replicates were analyzed. D - Histogram showing a distribution of number of putative eRNAs paired to genes in RPB1-52R and RPB1-25R cells.
Data matrix showing frequent SCNVs, significantly mutated genes (bold; Benjamini-Hochberg FDR 0.figf2Data matrix showing frequent SCNVs, significantly mutated genes (bold; Benjamini-Hochberg FDR 0.2 calculated with CHASM) and their associated pathway or family mutations discovered in cases with NPC analyzed by WES, targeted deep sequencing (TS) or SNP array (SNP-a). The frequencies of the alterations are plotted on the right. The colors and shapes denoting different types of somatic events are also applied in Figure 5a. Columns, examined cases; rows, genes; hot spot, identical mutations have been registered in COSMIC (see URLs); LOH, loss of heterozygosity. The asterisk indicates that this tumor was also subjected to SNP array profiling. PIK3CA amplification in C666 cells has been previously described.
Figure 5MYC down-regulates the m6A modification preferentially in certain MRGs, by up-regulating the demethylase ALKBH5. The m6A reader YTHDF3-mediated translation of MRGs SPI1 and PHF12 is attenuated as decreased m6A modification, and thus releases the inhibitory effect of MRGs on cell proliferation, thereby promoting cancer progression. The up-direction red arrows indicate high and down-direction red arrows indicate low.
Online lung microarray analysis revealed PIP as one of the Slug transcriptional targets mediating vascular cell proliferation in PF-PH.Figure 3(A) Heatmap of the 20 genes up-regulated in GSE24988 microarray and known to be Slug transcriptionnal target. (B) Venn Diagram showing the overlap between Slug targets implicated in cell proliferation and known to be extracellular. (C) Heatmap of the seven transcriptional targets of Slug known to promote proliferation and to be extracellular. Relative expression of Prolactin-induced protein (PIP) mRNA (D) normalized to GAPDH in PF and PF-PH patients. Relative expression of Prolactin-induced protein (PIP) expression normalized to GAPDH in PF and PF-PH patients. (F) Representative images of PIP immunohistochemistry showing its extracellular localization and quantification. Proliferation assay on healthy fibroblast (G) Pulmonary arterial EC (H) and on healthy pulmonary arterial SMC (I in the presence or absence of PIP protein. The number of samples per group for each experiment are included within each bar graph. Proliferation assay on healthy pulmonary arterial SMC J) in the presence or absence of PIP protein. The number of samples per group for each experiment are included within each bar graph.
a) Top: immunoblots for the indicated proteins from two independent experiments.figf1(a) Top: immunoblots for the indicated proteins from two independent experiments. Bottom: quantification of changes in total LRRK2 (n = 6 independent experiments). SH-SY5Y cells were untreated or incubated with 5 μM lactacystin (Lact), 10 mM 3MA or 20 mM NH4Cl and 100 μM leupeptin (NL) for 9.5 h. (b) Top: representative immunoblots for the indicated proteins from two independent experiments. Bottom: quantification of LRRK2 relative to that in untreated samples (n = 3 or 4 independent experiments). Brain slices from mouse cortex, midbrain or cerebellum were incubated without additions or with NL or 3MA for 2 h at 37 °C. (c,d) Homogenate (Hom) and lysosomal fractions (Lys) isolated from SH-SY5Y cells (c) or the indicated mouse brain regions (d), immunoblotted for the indicated proteins. (e) HEK293 cells expressing GFP-tagged LRRK2 and transduced with lentivirus control (Ctr) or carrying shRNA against LAMP-2A (L2A(−)), incubated or not with NL for 12 h and immunoblotted for the indicated proteins. (f) Immunoblots for LRRK2 and LAMP-2A of homogenates and lysosome-enriched fractions isolated from control fibroblasts (Ctr) or fibroblasts stably interfered for LAMP-2A (L2A(−)), maintained in the presence or absence of serum. Actin and LAMP-1 (L-1) are shown as loading controls of homogenate and lysosomes, respectively. All values are mean + s.e.m.; *P 0.05, unpaired t-test. Full-length blots and gels in Supplementary Figure 12.
Figure 3A, Cortical sections from rTg4510:Fluc-EGFP mice (right) and control littermates (left) stained for total human tau (HT7, magenta) and p-tau (AT8, yellow). Fluc-EGFP was detected by EGFP fluorescence (green), nuclei were labeled with DAPI (blue). Images on the right show higher magnification of the areas indicated by the boxes, with neuronal cell bodies outlined. Note the presence of EGFP foci (arrowheads) in the majority of neurons in rTg4510:Fluc-EGFP mice. Occasional foci of lower relative fluorescence intensity can be observed in control mice (arrow). B, Quantification of fluorescence intensity of EGFP foci, normalized to cytoplasmic EGFP fluorescence in cells with and without tau expression and p-tau pathology. N=4 rTg4510:Fluc-EGFP mice, 3 WT:Fluc-EGFP mice, 3 CaMKIIα-tTA:Fluc-EGFP mice and 3 tetO-tauP301L:Fluc-EGFP mice. WT:Fluc-EGFP, CaMKIIα-tTA:Fluc-EGFP and tetO-tauP301L:Fluc-EGFP littermates did not differ from each other and were pooled together as controls. One-way ANOVA with Tukey's multiple comparisons test. ANOVA: ****p < 0.0001. Pairwise comparisons between groups are indicated on the graph. C, Contour plot of fluorescence intensity distribution of EGFP foci in cells with and without tau expression and p-tau pathology. N = 174 tau- / p-tau- neurons; 178 tau+ / p-tau- neurons, and 69 tau+ / p-tau+ neurons. D, Quantification of the fraction of cells with EGFP foci among the cells with and without tau expression and p-tau pathology. N=4 rTg4510:Fluc-EGFP mice, 3 WT:Fluc-EGFP mice, 3 CaMKIIα-tTA:Fluc-EGFP mice and 3 tetO-tauP301L:Fluc-EGFP mice. WT:Fluc-EGFP, CaMKIIα-tTA:Fluc-EGFP and tetO-tauP301L:Fluc-EGFP littermates did not differ from each other and were pooled together as controls. One-way ANOVA with Tukey's multiple comparisons test. ANOVA: ****p < 0.0001. Pairwise comparisons between groups are indicated on the graph. E, F, Luciferase assay in rTg4510:Fluc-EGFP mice. E, Left, scheme of brain regions used for luciferase assay color-coded by their vulnerability to tau pathology. Right, specific luciferase activity measured in the indicated brain regions of 4-month-old and 16-month-old rTg4510:Fluc-EGFP mice normalized to control littermates. F, Fluc-EGFP protein quantity in the indicated brain regions of rTg4510:Fluc-EGFP mice measured by Western blot (for representative blots, see Fig. EV3B). Values are normalized to control littermates. N=6 rTg4510:Fluc-EGFP mice, 4 WT:Fluc-EGFP mice, 6 CaMKIIα-tTA:Fluc-EGFP mice and 1 tetO-tauP301L:Fluc-EGFP mouse at 4 months; 4-5 rTg4510:Fluc-EGFP mice, 5 WT:Fluc-EGFP mice, 3 CaMKIIα-tTA:Fluc-EGFP mice and 4 tetO-tauP301L:Fluc-EGFP mice at 16 months. WT:Fluc-EGFP, CaMKIIα-tTA:Fluc-EGFP and tetO-tauP301L:Fluc-EGFP littermates did not differ from each other and were pooled together as controls. Light-red and dark-red asterisks indicate comparisons to the corresponding littermate group (one-sample t-test), black asterisks indicate significant comparisons between corresponding brain regions of rTg4510:Fluc-EGFP mice at 4 vs. 16 months (two-tailed t-test).
Figure 7(A) Schematic diagram depicting the distinctive impacts of the functional interplay between EZH2 protein and p53 mRNA on progression of cancers expressing WT p53, no p53 or GOF mutated p53. MTase, methyltransferase. (B) Kaplan-Meier plots showing the association of EZH2 overexpression with biochemical recurrence of prostate cancer in patients from the TCGA cohort. Tumors were separated into two groups according to EZH2 expression levels (high or low) and three groups based on the mutation status of p53 (WT, loss or mutated). VCaP and C4-2 cells were treated with GSK-126, DZNep (5 µM for VCaP and 10 µM for C4-2), and EZH2 ASO, followed by western blots with indicated antibodies (C) Data shown as means±SD (n=6). Statistical significance was determined by two-tailed Student's t-test. *** P < 0.001. VCaP and C4-2 cells were treated with GSK-126, DZNep (5 µM for VCaP and 10 µM for C4-2), and EZH2 ASO, followed by MTT assay (D). Data shown as means±SD (n=6). Statistical significance was determined by two-tailed Student's t-test. *** P < 0.001. (E, F) Effect of Ezh2 ASOs on growth of murine prostate cancer allografts. Mice (n=10) bearing the Ptenpc-/-;Trp53pc-/- (E) and Ptenpc-/-;Trp53pc-/R172H allografts (F) were treated with control ASOs (50 mg kg-1) or Ezh2 ASOs at different doses (25 mg kg-1 and 50 mg kg-1). Tumor growth was measured twice a week for 3 weeks and the data are shown in the bottom panels. Each allograft was isolated by the end of ASO treatment and photographed (top panels). Data shown as means±SD (n=10). Statistical significance was determined by two-tailed Student's t-test. *** P < 0.001.
Lack of Sen1 produces extended read-through tRNA transcripts.Figure 5Cells of the indicated genotypes that carried or not the DRT5T dimeric tRNA construct (schematized on top) were grown either in the presence of the optimum concentration of adenine (left) or in the presence of a limiting concentration of adenine (right). Two independent clones of the same genotype (#1 and #2) were used. See text for details. 3F refers to the 3 Flag epitope tag at the C-terminus of Rpc37. Tetrad dissection was used to show that the double mutant sen1∆ rpc37-V189D is dead. Strand-specific RT-qPCR was used to quantify the levels of read-through transcripts (see Methods). The mean ± standard deviation from 4 biological replicates is represented here. P-values were obtained using the Wilcoxon-Mann Whitney statistical test. Northern blot analysis of the tRNA SPATRNAPRO.02 using an intron-specific probe (TCTAAACTCAGCATACAAGTGGGG). U5 snRNA was used as a loading control. Sequence of the ~350 nt-long read-through transcript at SPATRNAPRO.02. Residues in blue represent the sequence of the mature tRNA. Residues in red represent potential terminator sequences. Red arrows show the 3' end nucleotide of the read-through transcripts. The numbers indicate the number of times the sequenced transcripts terminated at the indicated position.