10 research outputs found
Characterization of TBCK.
<p>(<b>A</b>) Schematic comparison of TBCK homologs in <i>H. sapiens</i>, <i>M. musculus</i>, <i>D. rerio</i>, <i>D. melanogaster</i>, and <i>C. elegans</i>. Red, green and blue bars indicate possible kinase domain, TBC domain and rhodanese homology domain, respectively. (<b>B</b>) Expression of TBCK in mammalian cell lines. Total lysates from the indicated cells were resolved by SDS-PAGE and subjected to Western analysis. (<b>C</b>) Subcellular localization of TBCK. HEK293 cells grown on cover slides were immunostained with the antibodies as shown. DNA was visualized by DAPI. Bar, 10 μm.</p
Depletion of TBCK inhibits cell proliferation.
<p>HEK293 cells infected with the lentiviruses targeting to the various regions of TBCK mRNA (TBCK-RNAi-1 or 2) were subjected to Western blotting (<b>A</b>), cell counts (<b>B</b>), fluorescence microscopy (<b>C</b>) and MTT analyses (<b>D</b>). Bar, 200 µm. Lentivirus-infected cells were GFP-positive. Data are shown as the mean of three independent experiments ± SE (* <i>P</i><0.05, ** <i>P</i><0.01).</p
TBCK Influences Cell Proliferation, Cell Size and mTOR Signaling Pathway
<div><p>Mammalian target of rapamycin (mTOR) is a central regulator for both cell proliferation and cell growth; however, little is known about the regulation of mTOR expression at the transcriptional level. Here, we provide evidences that a conserved human protein TBCK (TBC1 domain containing kinase) is involved in the regulation of mTOR signaling pathway. Depletion of TBCK significantly inhibits cell proliferation, reduces cell size, and disrupts the organization of actin, but not microtubule. Knockdown of TBCK induces a significant decrease in the protein levels of components of mTOR complex (mTORC), and suppresses the activity of mTOR signaling, but not MAPK or PDK1/Akt pathway. Further results show that TBCK influences the expression of mTORC components at the transcriptional level. Thus, these data suggest that TBCK may play an important role in cell proliferation, cell growth and actin organization possibly by modulating mTOR pathway.</p></div
Depletion of TBCK influences mTOR signaling pathway.
<p>HEK293 cells were infected with the indicated lentivirus and subjected to Western blotting (<b>A</b>) and immunofluorescence analysis (<b>B</b>–<b>C</b>) with the antibodies as shown. GFP-positive signals indicate the cells infected by lentivirus. DNA was visualized by DAPI. Bar, 10 µm.</p
TBCK depletion reduces cell size.
<p>HEK293 cells were infected with the indicated lentivirus and subjected to fluorescence microscopy (A–B) and FACS analysis (C–D). (<b>A</b>–<b>B</b>) GFP-positive signals indicate the cells infected by lentivirus. Bar, 20 µm. The area measurements of infected cells were quantified by Image-Pro Plus 6.0 software. Data are shown as the mean of three independent experiments ± SE (** <i>P</i><0.01, n>200). (<b>C</b>–<b>D</b>) Representative histogram of flow cytometry shows the size distribution (FSC-H) of GFP positive cells that were stained with propidium iodide. The mean of three independent experiments ± SE is shown (* <i>P</i><0.05, n>100,000).</p
TBCK has no significant effect on MAPK and PDK1/AKT pathway.
<p>(<b>A</b>–<b>B</b>) HEK293 cells infected with the indicated lentivirus were subjected to Western blotting with the antibodies as shown.</p
TBCK plays a role in actin organization.
<p>HEK293 cells infected with the indicated lentivirus were subjected to immunofluorescence staining with phalloidin-TRITC (<b>A</b>) or anti-α-tubulin antibody (<b>C</b>). Lentivirus-infected cells were GFP-positive. DNA was visualized by DAPI. Bar, 10 µm. Single panel image of phalloidin-TRITC signal (<b>B</b>) or α-tubulin (<b>D</b>) is showed in rainbow palettes.</p
TBCK influences the protein level of mTOR components and the activities of mTOR complexes.
<p>(<b>A</b>–<b>B</b>) HEK293 cells infected with the indicated lentivirus were subjected to immunoblotting with the antibodies as shown.</p
Depletion of TBCK represses the mRNA level of mTOR complexes components.
<p>Total RNAs from HEK293 cells infected with the indicated lentivirus were analyzed by quantitative RT-PCR with the targeted genes as shown. Error bars indicate SE (** <i>P</i><0.01).</p
Enhanced Photothermocatalytic Synergetic Activity Toward Gaseous Benzene for Mo+C-Codoped Titanate Nanobelts
In the present paper, a simple and facile method is proposed
to
synthesize Mo+C-codoped titanate (H<sub>2</sub>Ti<sub>5</sub>O<sub>11</sub>·3H<sub>2</sub>O) nanobelts by using hydrothermal method
together with sol–gel technique, and various Mo-doped and Mo+C-codoped
titanate nanobelts are realized by controlling the precursor’s
contents. It is found that the photocatalytic activity of titanate
nanobelts toward gaseous benzene will be greatly enhanced by Mo+C-codoping
if comparing with that of the pure titanate nanobelts, Mo-doped titanate
nanobelts, and C-doped titanate nanobelts, and the optimal Mo doping
content is confirmed to be 0.33 wt % for Mo/Ti ratio. The reason lies
in the fact that by Mo+C-codoping (Mo/Ti ratio of 0.33 wt %, C/Ti
ratio of 0.33 wt %), the band gap of titanate nanobelts will be narrowed
and the allowed paths for photoexcitation will be increased with the
generation of impurity energy level, which results in an obvious red-shift
in their UV–vis absorption spectra and the increased utilization
of the solar energy, and thus enhances their photocatalytic activity.
On the basis of the above experimental results, the current photocatalytic
mechanism is proposed