EMBO/sd-character-level-ner huggingface dataset:
Life-long TFEB treatment improves behavioral performance and synaptic functionMorris water maze test of groups of 4-month-old WT or Tau mice without or with TFEB (+TFEB) treatment, showing longer latency (time to find hidden platform) in Tau mice compared to WT controls at any given day of training.Figure 5A Morris water maze test of groups of 4−month−old WT or Tau mice without or with TFEB (+TFEB) treatment, showing longer latency (time to find hidden platform) in Tau mice compared to WT controls at any given day of training. TFEB has no effect on WT mice, but significantly reduced the latency in Tau mice. n = 14/group. ***P < 0.001 (2 way ANOVA). B Number of platform crossings in non‐target quadrants (NTQ) vs. the target quadrant (TQ). Values of the three NTQs were combined and averaged. n.s.: non‐significant. *P = 0.047 and ***P = 0.0005 for TQ of WT vs Tau and Tau vs Tau + TFEB, respectively (Student's t‐test). Each bar represents average ± s.e.m. C Slope of field excitatory postsynaptic potential (fEPSP) in response to theta-burst stimulation delivered to the Schaffer collateral pathway from WT mice (n = 10 recordings from 6 mice), Tau mice (n = 19 recordings from 6 mice) or Tau mice with TFEB injection (Tau + TFEB; n = 8 recordings from 6 mice). Insets: example fEPSP traces taken before (upper) or after (lower) stimulation from WT, Tau or Tau + TFEB mice. Calibration: 1 mV, 5 msec. D Quantification of average fEPSP slope in the last 10 minutes demonstrating severely impaired LTP in Tau mice (WT vs. Tau) and significant improvement by TFEB treatment (Tau vs. Tau + TFEB). Each bar represents average ± s.e.m. **P < 0.01 (2 way ANOVA with Bonferroni post hoc). E Cytoscape‐generated networks representing genes involved in synaptic function. A majority of downregulated genes (blue dots) are found in the synaptic network when comparing transcription profiles of Taumice with wild‐type controls (Tau vs. WT), while a majority of upregulated genes (red dots) are found when comparing TFEB‐injected with uninjected Taumice (Tau + TFEB vs. Tau). n = 4 mice/genotype/treatment group.
Figure 5(A) Clusters of genes, whose expression changes towards the hepatocyte fate, upon removal of R-spondin 1 and noggin from the organoid culture media (Table EV5). The number of genes and the names of selected genes in each cluster are indicated. Gene expression levels (log2 scale) are shown for non-differentiated (N) and differentiated (D) gallbladder-derived organoids (GO), gallbladder tissue (GT) and liver tissue (LT). Upreg, upregulated; Downreg, downregulated. (B) Expression levels of selected genes from the clusters of panel A. GO, gallbladder organoid; N, non-differentiated; D, differentiated; G, gallbladder; L, liver; T, tissue. (C) Removal of R-spondin 1 and noggin from the media induces expression of red fluorescent protein (RFP) in a subset of gallbladder organoid cells. A transgene expressing tamoxifen-inducible Cre is driven by the endogenous Alb promoter; a second transgene expresses RFP after Cre-mediated rearrangement. Scale bar: 200 m. (D) Glycogen accumulation upon organoid differentiation, as revealed by periodic acid-Schiff (PAS) staining (red). Scale bar: 25 m.
Acs2p Controls ATG7 Transcription through Epigenetic Histone Acetylation (A) ATG7 mRNA levels by quantitative reverse-transcriptase PCR (RT-qPCR) of wild-type and Δach1 cells aged to day 3 (see also ).Figure 5(A) ATG7 mRNA levels by quantitative reverse-transcriptase PCR (RT-qPCR) of wild-type and Δach1 cells aged to day 3 (see also Figure S5). Rel. mRNA levels are expressed as ratios of 18S rRNA normalized to wild-type cells by ΔΔCt-method. Data represent means ± SEM (n = 8).(B) Representative immunoblot analysis from wild-type (WT) and ACH1-deleted (Δach1) yeast expressing chromosomally tagged ATG7 by C-terminal 6HA fusion aged to indicated time points. Blots were probed with anti-HA and anti-GAPDH (loading control) antibodies. (C and D) Representative immunoblots (C) and densitometric quantification (D) of whole-cell acid extracts of wild-type and Δach1 cells combined with or without knockdown of ACS2 (tet-ACS2). Cells were chronologically aged to day 3 in the presence of 1 ng/ml doxycycline. Blots were probed with antibodies against total histone H3 (loading control) or H3 acetylated lysines (K9Ac, K14Ac, K18Ac). Data represent means ± SEM (n = 8). (C and D) Representative immunoblots (C) and densitometric quantification (D) of whole-cell acid extracts of wild-type and Δach1 cells combined with or without knockdown of ACS2 (tet-ACS2). Cells were chronologically aged to day 3 in the presence of 1 ng/ml doxycycline. Blots were probed with antibodies against total Histone H3 (loading control) or H3 acetylated lysines (K9Ac, K14Ac, K18Ac). Data represent means ± SEM (n = 8). (E and F) ATG7 mRNA levels (E) by RT-qPCR and representative immunoblot analysis (F) of wild-type and Δach1 cells with or without knockdown of ACS2 (tet-ACS2) as in (C) and (E) aged to day 3. Rel. mRNA levels (E) are expressed as ratios to 18S rRNA normalized to wild-type cells by ΔΔCt method. Data represent means ± SEM (n = 7-8). (E and F) ATG7 mRNA levels (E) by RT-qPCR and representative immunoblot analysis (F) of wild-type and Δach1 cells with or without knockdown of ACS2 (tet-ACS2) as in (C) and (E) aged to day 3. Rel. mRNA levels (E) are expressed as ratios to 18S rRNA normalized to wild-type cells by ΔΔCt method. Data represent means ± SEM (n = 7-8). (G and H) Representative immunoblot (H) and densitometric quantification (G) of histone H3 wild-type (H3-wt/wt) and H3 mutated (H3-K14,18Q/-K14,18R) strains carrying the GFP-Atg8p fusion to calculate "free-GFP/GFP-Atg8p Ratio" indicative of autophagic flux.
Figure 2A HEK293 cells transfected with empty vector (lanes 1, 6), ATZ-HA (2, 7), FAM134B-V5 (3 and 8), FAM134B-V5 and ATZ-HA (4, 9), or FAM134BLIR-V5 and ATZ-HA (5, 10), incubated for 12 h with 50 nM BafA1 and then treated with the cross-linker DSP before lysis as described in Methods. Lanes 1-5, WB of the total cell extract (TCE); lanes 6-10, WB of antiV5 immunoprecipites to isolate complexes containing ectopically expressed FAM134B or FAM134BLIR. The membranes were probed with antiV5 (upper panels), antiCNX, antiHA and antiLC3 antibodies. B Quantification of LC3 co-precipitating with FAM134B-V5 (A, lanes 8 and 9). Mean ± SEM, n=3, unpaired two-tailed t-test, * P<0.05. WT MEF. Scale bar: 10 μm. D Ectopic expression of FAM134BLIR in WT MEF inhibits ATZ delivery to endolysosomes. Scale bar: 10 μm. E, F Distribution of gold-labeled ATZ-HA by IEM in BafA1 treated WT MEF and WT MEF overexpressing FAM134BLIR, respectively. EV, ER-derived vesicles, red arrowheads; EL, endolysosome. G Quantification of ATZ-gold density of E, F (n=75 and 79 EL, respectively). Unpaired two-tailed t-test, **** P <0.0001. H Max projection of the same cell as in D after deconvolution. Insets show orthogonal section of select regions. Scale bar: 10 μm. I Decay of ATZ polymers (CHX chase, upper panel) immunoisolated with the polymer-specific 2C1 antibody (visualized with antiHA in WB) in HEK293 cells mock-transfected (lanes 1-4) or expressing FAM134BLIR-V5 (lanes 5-8). Middle panel, expression of FAM134BLIR-V5 assessed by WB; lower panel, loading control. (J) Quantification of (I) (Mean ± SEM, n=3, unpaired two-tailed t-test, * P<0.05). K Flow cytometry analysis of ATZ-HA polymer levels in MEFs mock treated, exposed to BafA1, and co-expressing FAM134LIR. MFI: mean fluorescence intensity (Mean ± SEM, n=5, unpaired two-tailed t-test, ns P>0.05, ** P<0.01, *** P <0.001).
Figure 3Knock-down of βarr1 leads to an increase in agonist-induced ERK1/2 phosphorylation for B2RΔG368 and B2RΔI374 but a decrease for B2RL370T and B2RΔG368+L370T mutants. HEK-293 cells expressing the indicated B2R mutants in presence and absence of βarr1 knock-down were stimulated with indicated doses of bradykinin (Brady) for 10 min followed by detection of phosphorylated ERK1/2 using Western blotting. Densitometry-based quantification of data (mean±SEM) from three independent experiments (five for B2RΔG368 and B2RΔI374) is presented as bar graphs in the lower panels. Data are normalized with respect to maximal signal under control condition (treated as 100%), and analyzed using Two-way-ANOVA with Bonferroni multiple comparisons test (*p <0.05; **p <0.01; ***p<0.001).
Figure 1 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)  B mRNA expression in MACS-isolated Lin-Sca1+ progenitor cells from female mouse subcutaneous fat, differentiated in the presence of 100 nM Rosi or vehicle for the indicated time, as determined by qRT-PCR (n=4) ****P=1*10-10 (Day 1 and 2), **P=0.001, in 2x2 ANOVA with Bonferroni posttests (Rosi vs. vehicle)  C mRNA expression in female Lin-Sca1+ cells, differentiated in the presence of 100 nM Rosi or vehicle for 8 days, as determined by qRT-PCR (n=3) t-test Cited4-/- vs. Cited4+/+ (Rosi) *P=0.013 (Ucp1), **P=0.004 (Cpt1b), *P=0.026 (Dio2)  D-F mRNA expression in primary SVF cells from human subcutaneous fat, differentiated in the presence of 100 nM Rosi (D) or vehicle (D-F), as determined by qRT-PCR at the indicated time points (n=5 patients). ♀/♂ represent individual data (D) ****P=3*10-5 (Day 2), ****P=3*10-6 (Day 6), ****P=4*10-9 (Day 10), ****P=3*10-7 (Day 14), in 2x2 ANOVA with Bonferroni posttests (Rosi vs. vehicle). (E, F) Pearson correlation coefficient (r) and P-value are shown  G mRNA expression in primary SVF cells from human female subcutaneous fat transfected with the indicated siRNA prior to differentiation in the presence of 100 nM Rosi for 9 days, as determined by qRT-PCR (n=3) ***P=0.0002 (CITED4), **P=0.002 (UCP1), *P=0.02 (UCP1), *P=0.035/0.026 (PPARG), ***P=0.0006 (SLC2A4), **P=0.002 (ADIPOQ) in one-way ANOVA with Tukey posttests (vs. siCtrl)
a) Helical wheel representation of the predicted N-terminal amphipathic helix (amino acids 4–26) of mouse Atg3 (wheel generated in Heliquest).figf5(b) Analysis of reaction efficiency for point mutants of the amphipathic helix. GL1 lipidation reactions were run on 400 nm and sonicated liposomes (30 mol% DOPE) with the indicated Atg3 mutants. The extent of GL1-PE formation (top gel) was determined from densitometry and is plotted as percentage of total GL1 (bar graph). The ability to form the Atg3-GL1 conjugate was also assessed for each mutant (bottom immunoblot). Note that this conjugate forms in every case, indicating that no mutant inhibited a step upstream in the reaction. Further, this conjugate specifically accumulates in reactions where lipidation is impaired. In both panels, the samples were prepared at the same time from the same materials and run simultaneously on multiple gels, as indicated by the lines. (c) Curvature dependence of K11L mutant binding to liposomes compared with wild-type Atg3. The binding reaction was performed as in Fig. 4b on 30 DOPE% liposomes. F, float-up; T, total input. (d) Lys 11 represents a tunable node through which curvature dependence of the lipidation reaction can be controlled. Lipidation assays were run as in b. Total GL1 lipidation was assessed from reactions with wild-type Atg3 or mutant forms of Atg3 in which position Lys 11 was changed as indicated. Lipidation efficiency on low PE/low curvature liposomes (30% PE, 400 nm) increases with increasing hydrophobicity of the new amino acid, while liposomes with high PE (55%) or high curvature (sonicated) are good substrates for all active forms of Atg3. Left: Coomassie gels of individual lipidation reactions. Right: densitometry of Coomassie-stained gels revealed the ratio of GL1-PE to the sum of all three GL1 species as in Fig. 2. Light grey bars are sonicated liposomes; dark grey bars are 400 nm extruded liposomes. For b,c, P values were calculated between sonicated and 400 nm samples of the same mutant. For d, P values represent comparison with WT protein on the same liposome size and composition. For all panels, n = 3 independent experiments, error bars represent standard deviation and **P 0.05; *P 0.01. Uncropped gels in Supplementary Fig. 7.
Figure 4A Expression of murine ERVs in blood of WT and G1 Tert-/- mice. n = 5 mice per group. B Expression of MMERGLN_LTR and MMERGLN-int in livers of WT and G1 Tert-/- mice treated with ENU or saline (control). n = 3 mice per group. C C Enriched biological processes (BP) for WT versus G1 Tert-/- mice. GO was analysed for upregulated genes in WT mice compared to G1 Tert-/- mice under control or ENU treatment. D Heat map showing log2 fold change differences of interferon-related genes expression between ENU treatment and control (saline) in WT and G1 Tert-/- mice by RNA-seq. n = 3 mice per group. E Heat map showing log2 fold change differences of cytokine/chemokine between ENU treatment and control (saline) in WT and G1 Tert-/- mice by cytokine/chemokine array. n = 5 mice per group. F F ELISA of CXCL10 in WT and G1 Tert-/- mice treated with poly(I:C) or saline (control). n = 5 mice per group.
Losartan treatment delays RDEB fibrosis progression in vivo.Figure 1Dorsal and palmar view of the right forepaw of the C7-hypomorphic mice. At the start of treatment, the mice were on average 5.5 weeks old. The treated group received 0.6 g losartan per liter drinking water. Shown are photographs of two untreated mice and two mice receiving losartan at the start, after 4 weeks, and at the end of the experiment after 7 weeks. Note the fibrosis-driven loss and fusion of digits with time; the arrows indicate digit fusion. Forepaws viewed dorsally or palmary after 7 weeks of treatment ± losartan. Shown is a good responder of losartan treatment and one mouse from the control group with rapid mutilation rate. Bar graph of forepaw digit length in age-matched untreated (blue) vs. losartan-treated C7-hypomorphic mice (red) after 2, 4, and 7 weeks of treatment. The length of digits at the start of the experiment was set to 100%; the quantification procedure is described in detail in the Materials and Methods section. Losartan very potently inhibited the reduction of digit length. Values represent mean ± S.D. Due to the inherent heterogeneity in disease progression, equal variance could not be expected and statistical significance was therefore analyzed by the unpaired t-test with Welch's correction; for 2 weeks, **P = 0.0021, and for 4 and 7 weeks, ***P < 0.001 (n = 14 per group).
Figure 5 (A) Representative immunoblots of LETM1, DRP1 total protein levels and activating phosphorylation (DRP1S616) in HeLa cells treated with either NT or LETM1 siR1, siR2 or siR3. GADPH is shown as a loading control. Immunoblot signals were quantified using ImageJ; data are displayed as means ± SEM of n=4 experiments except for siR3 DRP1S616 (n=2)  (B) One hour 35S pulse-labeling of mitochondrial proteins in HeLa cells silenced for LETM1 (siR1) or DRP1 (siD), or co-silenced for LETM1 and DRP1 (siR1D) for 144 h. Coomassie stained gels are used as loading controls and polypeptide assignments are indicated to the right  (C) Immunoblotting of cell lysates prepared as (B) with antibodies to respiratory chain subunits NDUFB8 and COXII, and GAPDH as loading control. Means ± SEM of n=3 experiments  (D) Confocal analysis of HeLa cells silenced for LETM1 (siR1), or for DRP1 (siD), or co-silenced with both siR1 and siD for 96 h. Mitochondrial network, mtDNA and cell nuclei were immunostained with anti-TOM20 (red) anti-DNA (green), and DAPI (blue), respectively. Scale bars are 6 µm in the main images and 3 µm in offset merged magnifications
Figure 8Wnt-induced deubiquitination and stabilization of FoxM1 abolishes the inhibitory effect of ICAT leading to transactivation of β-catenin hence promoting cellproliferation.
Figure 3(A) Growth curves of MCF10A cells exposed to oxidative stress (200μM H2O2) relative to control cells (n=3). Two-way ANOVA was used to test for significance. (B) Northern blot of MCF10A cells expressing a control short-hairpin RNA or a hairpin targeting tRNATyrGUA. (C) Western blot of MCF10A expressing a control short-hairpin RNA or a hairpin targeting the tyrosine-tRNA synthetase, YARS (red arrow). HSC70 was used as a loading control. (D) Growth curves of MCF10A cells expressing RNAi targeting mature tRNATyrGUA or YARS relative to cells expressing a control hairpin (n=3). Two-way ANOVA was used to test for significance. (E) Cell growth of MCF10A cells transiently transfected with a tRNATyrGUA overexpression vector relative to an empty control vector (n=3). A one-tailed Mann-Whitney test was used to test for significance at day 3.
Figure 4(A) HeLa cells were transfected with siControl or siCyclin B2. At 36 h after transfection, cells were collected and lysed. The samples were resolved by SDS-PAGE and analyzed by Western blotting using anti-Cyclin B2 antibody, anti-Cyclin B1 antibody, and anti-tubulin antibody (B) Representative stills illustrating mitotic progression in mCherry-H2B expressed cells treated with Control siRNA or Cyclin B2 siRNA. Images were acquired at the indicated time points after the start of NEBD. The arrow indicates lagging chromosomes (C) Bar graph illustrating the percentage of cells with normal anaphase or with anaphase lagging chromosomes in cells treated as in B. Bars are means ±SD (≥30 cells from three independent experiments) (D) Scatter graph of the time from NEBD to the beginning of anaphase in cells treated as in B (n = 30 cells) (E) Representative stills illustrating mitotic progression in mCherry-H2B expressed cells depleted of endogenous Cyclin B2 and rescued with GFP-Cyclin B2-WT or GFP-Cyclin B2-4A. Images were acquired at the indicated time points after the start of NEBD (F) Bar graph illustrating the percentage of cells with normal anaphase and cells with anaphase lagging chromosomes in cells treated as in E. Bars are means ±SD (≥30 cells from three independent experiments)
a) COS-7 cells expressing EGFP-tagged wild-type (Gln23; Q23) or mutant (Gln74; Q74) huntingtin exon 1 (green) for 48 h stained for endogenous mTOR (red) show mTOR in mutant aggregates.figf1a) COS-7 cells expressing EGFP-tagged wild-type (Gln23; Q23) or mutant (Gln74; Q74) huntingtin exon 1 (green) for 48 h stained for endogenous mTOR (red) show mTOR in mutant aggregates. (b) Double-labeling immunohistochemistry of huntingtin (EM48; green) and mTOR (red) in brain tissue of mice expressing mutant huntingtin. (c) Immunohistochemistry of brain tissue from individuals with grade III Huntington disease or unaffected controls using antibody to mTOR. (d) Western blot of lysates from stable inducible PC12 cells 48 h after expression of wild-type (Gln23; Q23) or mutant (Gln74; Q74) huntingtin showing mTOR in the stacking gel (mTOR insoluble) in cells expressing mutant huntingtin. (e) High-molecular-mass aggregates of mTOR in both PC12 (i) and COS-7 (ii) cells expressing mutant (Gln74; Q74) huntingtin. Blots of cell lysates from PC12 cells were probed with antibodies to phosphorylated mTOR (Ser2448; p-mTOR; iii) and to GFP (iv); both showed insoluble material in the stack. (f) Cell lysates from stable inducible PC12 cells and transiently transfected COS-7 cells expressing wild-type (Gln23; Q23) or mutant (Gln74; Q74) huntingtin for 48 h were immunoprecipitated using antibody to GFP and immunoblotted with antibody to mTOR. Uninduced PC12 cells with mutant huntingtin (UI) and untransfected COS-7 cells (UT) were controls. (g) Brain lysates from mice expressing mutant huntingtin (Tg) or control (Cont) littermates were immunoprecipitated with antibody to mTOR and immunodetected with an antibody recognizing the polyglutamine tract (IC2). Htt, huntingtin
Figure 1qRT-PCR of mGPDH, myogenin and myosin heavy chain (MyHC) levels during C2C12 myocyte differentiation immunoblot of mGPDH, myogenin and myosin heavy chain (MyHC) levels during C2C12 myocyte differentiation. Quantification represents the levels of indicated protein normalized to β-actin Immunoblot of mGPDH, voltage-dependent anion channel (VDAC) and cytochrome c (Cyt C) levels in mitochondrial lysate during C2C12 myocyte differentiation. Quantification represents the levels of indicated protein normalized to COX IV Activity assay of mGPDH at days 0 and 7 after C2C12 myocyte differentiation Representative images of MyHC immunofluorescence (E) of C2C12 myocyte transfected with the siRNA or the overexpression plasmid for mGPD C2C12 myocyte transfected with the siRNA or the overexpression plasmid for mGPDH; the fusion index (F) and the distribution of nuclei per myotube (G) were calculated at day 5 after differentiation. Immunoblot of mGPDH, myogenin and MyHC in C2C12 myocytes transfected with siRNA targeting mGPDH. Quantificatio represents the levels of indicated protein normalized to β-actin at indicated day after differentiation myogenin and MyHC in C2C12 myocytes transfected with siRNA targeting mGPDH. Quantificatio represents the levels of indicated protein normalized to β-actin at indicated day after differentiation qRT-PCR analysis of mGPDH, myogenin and MyHC in C2C12 myocytes transfected with the siRN for mGPDH at day 4 after differentiation qRT-PCR analysis of mGPDH, myogenin and MyHC in C2C12 myocytes transfected wit overexpression plasmid for mGPDH at day 4 after differentiation
Figure 3A Schematic representation of tiTELs. An nmt1 promoter (Pnmt1) was inserted upstream of the natural TERRA TSS. Telomeric repeats are in black and the subtelomeric tract in grey. A cassette conferring resistance to nourseothricin (natr) was used to select for positively transformed cells. The inserted cassette generates a novel HindIII restriction site, while the endogenous ApaI restriction site close to the telomeric tract is disrupted upon integration. Probes and oligonucleotides used for northern blot, Southern blot and RT-PCR analyses are indicated. The sketch is not to scale. B Genomic DNA from wt cells (CAF13) and cells carrying two tiTELs (CAF110) maintained in YES medium was digested with HindIII and Southern blot-hybridized consecutively with the indicated probes. RET refers to telomeric sequences (tiTELs or natural telomeres, nTELs) retained in the upper part of the gel; REL refers to telomeric sequences released in the lower part of the gel. The black arrow points to the natural nmt1+ sequence on chromosome III. Marker molecular weights are on the left in kilobases. C 5' RACE analysis of the tiTERRA transcription start site using total RNA from cells carrying two tiTELs (CAF110) maintained in YES medium. Nucleotides boxed in blue overlap with the 5' UTR of the nmt1+ transcript while the ones in green correspond to subtelomeric sequences. tiTERRA and TERRA transcription start sites (TSS) are indicated. D qRT-PCR analysis of tiTERRA in cells carrying no tiTEL (CAF13), one tiTEL (CAF545) or two tiTELs (CAF110) cultured in EMM medium in presence or absence of Thiamine (THI; repressed and induced condition, respectively) for 24 h. RNA was reverse transcribed with oC and cDNA was amplified with oF3 and oR3 oligonucleotides to detect only tiTERRA (graph on the left) or with oF1 and oR1 oligonucleotides to detect simultaneously tiTERRA and natural TERRA (on the right). Values are normalized to ACT1 mRNA and expressed as fold increase over CAF545 in uninduced conditions (THI+). Bars and error bars are averages and SD from 3 independent experiments. * P < 0.05, ** P < 0.01 (relative to CAF545 THI+; two-tailed Student's t test). E Northern blot analysis of total RNA from wt cells and cells carrying two tiTELs (CAF110) treated with combinations of THI and Trichostatin A (TSA) for 24 h. Two identical membranes were hybridized in parallel with oligonucleotides corresponding to C-rich (oC) or G-rich (oG) telomeric repeats. To confirm similar hybridization efficiencies of the two oligonucleotides, northern blot membranes were simultaneously hybridized and exposed along with blots of digested genomic DNA (upper right insets). After signal detection, RNAmembranes were stripped and re-hybridized with U6 probes to assure equal loading. Marker molecular weights are on the left in kilobases.