535 research outputs found
The Switch from NF-YAl to NF-YAs Isoform Impairs Myotubes Formation
NF-YA, the regulatory subunit of the trimeric transcription factor (TF) NF-Y, is regulated by alternative splicing (AS) generating two major isoforms, "long" (NF-YAl) and "short" (NF-YAs). Muscle cells express NF-YAl. We ablated exon 3 in mouse C2C12 cells by a four-guide CRISPR/Cas9n strategy, obtaining clones expressing exclusively NF-YAs (C2-YAl-KO). C2-YAl-KO cells grow normally, but are unable to differentiate. Myogenin and-to a lesser extent, MyoD- levels are substantially lower in C2-YAl-KO, before and after differentiation. Expression of the fusogenic Myomaker and Myomixer genes, crucial for the early phases of the process, is not induced. Myomaker and Myomixer promoters are bound by MyoD and Myogenin, and Myogenin overexpression induces their expression in C2-YAl-KO. NF-Y inactivation reduces MyoD and Myogenin, but not directly: the Myogenin promoter is CCAAT-less, and the canonical CCAAT of the MyoD promoter is not bound by NF-Y in vivo. We propose that NF-YAl, but not NF-YAs, maintains muscle commitment by indirectly regulating Myogenin and MyoD expression in C2C12 cells. These experiments are the first genetic evidence that the two NF-YA isoforms have functionally distinct roles
Role of the lung resistance related protein (LRP) in the drug sensitivity of cultured tumor cells
Relativistic effects and two-body currents in using out-of-plane detection
Measurements of the reaction were performed
using an 800-MeV polarized electron beam at the MIT-Bates Linear Accelerator
and with the out-of-plane magnetic spectrometers (OOPS). The
longitudinal-transverse, and , and the
transverse-transverse, , interference responses at a missing momentum
of 210 MeV/c were simultaneously extracted in the dip region at Q=0.15
(GeV/c). On comparison to models of deuteron electrodisintegration, the
data clearly reveal strong effects of relativity and final-state interactions,
and the importance of the two-body meson-exchange currents and isobar
configurations. We demonstrate that these effects can be disentangled and
studied by extracting the interference response functions using the novel
out-of-plane technique.Comment: 4 pages, 4 figures, and submitted to PRL for publicatio
Different vimentin expression in two clones derived from a human colocarcinoma cell line (LoVo) showing different sensitivity to doxorubicin.
We selected two clones, isolated from the human colocarcinoma cell line LoVo, showing a sensitivity to doxorubicin similar to (LoVo clone 5) or three times lower than (LoVo clone 7) the parental cell line. Since vimentin was atypically expressed in a human breast carcinoma cell line made resistant to doxorubicin, we looked at vimentin expression in these two clones with spontaneously different sensitivity to the drug. For comparison we used the parental cell line LoVo WT and LoVo/DX made resistant pharmacologically. mRNA for vimentin was undetectable by Northern blot analysis in LoVo WT and in LoVo clone 5, while expression of this gene was high in LoVo clone 7 and in LoVo/DX. This increase in mRNA levels was not related to an amplification of DNA, as suggested by Southern blot analysis. Immunofluorescence and immunocytochemistry findings confirmed, at protein level, the mRNA data. In LoVo clones 5 and 7, there were respectively 8.6% and 71% vimentin-positive cells, although the two clones showed similar expression of multidrug resistance gene 1 (mdr-1) and accumulated intracellular doxorubicin at similar levels. Similarly, drug efflux was the same for both clones. Our results show for the first time that cells resistant to doxorubicin express vimentin independently of the mdr glycoprotein. However when cells from clone 5 were transfected with human vimentin cDNA, they did not become resistant, indicating that vimentin can be considered as a marker of resistance in these cells but does not give rise to a resistant phenotype by itself
Characterization of a clonal human colon adenocarcinoma line intrinsically resistant to doxorubicin.
Intrinsic low-level resistance to anti-cancer drugs is a major problem in the treatment of gastrointestinal malignancies. To address the problem presented by intrinsically resistant tumours, we have isolated two monoclonal lines from LoVo human colon adenocarcinoma cells: LoVo/C7, which is intrinsically resistant to doxorubicin (DOX); and LoVo/C5, which shows the same resistance index for DOX as the mixed parental cell population. For comparison, we have included in the study a LoVo-resistant line selected by continuous exposure to DOX and expressing a typical multidrug resistant (MDR) phenotype. In these cell lines we have studied the expression and/or activity of a number of proteins, including P-glycoprotein 170 (P-gp), multidrug resistance-associated protein (MRP), lung resistance-related protein (LRP), glutathione (GSH)-dependent enzymes and protein kinase C (PKC) isoforms, which have been implicated in anti-cancer drug resistance. Intracellular DOX distribution has been assessed by confocal microscopy. The results of the present study indicate that resistance in LoVo/C7 cells cannot be attributed to alterations in P-gp, LRP or GSH/GSH-dependent enzyme levels. Increased expression of MRP, accompanied by alterations in the subcellular distribution of DOX, has been observed in LoVo/C7 cells; changes in PKC isoform pattern have been detected in both intrinsically and pharmacologically resistant cells
WARP: a WIMP double phase Argon detector
The WARP programme for dark matter search with a double phase argon detector
is presented. In such a detector both excitation and ionization produced by an
impinging particle are evaluated by the contemporary measurement of primary
scintillation and secondary (proportional) light signal, this latter being
produced by extracting and accelerating ionization electrons in the gas phase.
The proposed technique, verified on a 2.3 liters prototype, could be used to
efficiently discriminate nuclear recoils, induced by WIMP's interactions, and
measure their energy spectrum. An overview of the 2.3 liters results and of the
proposed 100 liters detector is shown.Comment: Proceeding for IDM200
Speech listening entails neural encoding of invisible articulatory features
Speech processing entails a complex interplay between bottom-up and top-down computations. The former is reflected in the neural entrainment to the quasi-rhythmic properties of speech acoustics while the latter is supposed to guide the selection of the most relevant input subspace. Top-down signals are believed to originate mainly from motor regions, yet similar activities have been shown to tune attentional cycles also for simpler, non-speech stimuli. Here we examined whether, during speech listening, the brain reconstructs articulatory patterns associated to speech production. We measured electroencephalographic (EEG) data while participants listened to sentences during the production of which articulatory kinematics of lips, jaws and tongue were also recorded (via Electro-Magnetic Articulography, EMA). We captured the patterns of articulatory coordination through Principal Component Analysis (PCA) and used Partial Information Decomposition (PID) to identify whether the speech envelope and each of the kinematic components provided unique, synergistic and/or redundant information regarding the EEG signals. Interestingly, tongue movements contain both unique as well as synergistic information with the envelope that are encoded in the listener's brain activity. This demonstrates that during speech listening the brain retrieves highly specific and unique motor information that is never accessible through vision, thus leveraging audio-motor maps that arise most likely from the acquisition of speech production during development
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