44 research outputs found
Inverse correlation between the concentration of HGF in culture and production of HGF by HSCs.
<p>(<b>A</b>) Real-time PCR for HGF mRNA shows a dose-dependent decrease in HGF production by HSC-T6 cells in response to increasing concentrations of HGF in culture (*P<0.05; **P<0.01). (<b>B</b>) HGF protein production by HSC-T6 cells decreases in response to increased HGF, as measured by WB. Actin represents loading control. (<b>C</b>) Densitometry analysis on representative WB shown in (B).</p
Genotype and number of mice used in experiments.
<p>Genotype and number of mice used in experiments.</p
Liver regeneration stimulated by CCl4 depletes HGF mRNA and protein in HGF KO mice.
<p>(<b>A</b>) CCl4 treatment following genomic recombination further decreases full-length HGF mRNA, as assessed by real-time PCR. (<b>B</b>) WB shows decreased HGF expression in livers of HGF KO mice treated with CCl4 in combination with p(I):p(C), as compared to controls or those treated with p(I):p(C) only. Ponceau represents loading control.</p
Recombination occurs in all hepatic cell populations, including those that produce HGF.
<p>(<b>A</b>) Separation of hepatic cell populations from HGF<sup>ex.5 flox</sup>;Mx1-cre mice into hepatocytes and NPCs shows recombination in both after p(I):p(C) treatment as compared to controls. (<b>B</b>) RT-PCR shows persistence of unrecombined HGF mRNA in both hepatocytes and NPCs. (<b>C</b>) Real-time PCR for full-length HGF mRNA shows a decrease in HGF in the NPC fraction after p(I):p(C) treatment. (<b>D</b>) WB for HGF in hepatocytes and NPCs shows that the amount of HGF is unchanged in KOs compared to controls, and is found mainly in the NPC fraction. Ponceau represents loading control.</p
Persistence of unrecombined HGF mRNA and protein in the livers of HGF<sup>ex.5 flox</sup>; Cre<sup>+/−</sup> mice after genomic recombination.
<p>(<b>A</b>) Schematic of the targeting strategy for conditional inactivation of the gene for HGF (top). Cre-mediated excision of the floxed HGF allele (middle) leads to the generation of a recombined allele (bottom) lacking exon 5. (<b>B</b>) Successful genomic deletion of HGF exon 5 after induction of recombination, as shown by PCR. Top - HGF<sup>ex.5 flox</sup>;Cre-ER<sup>T</sup> mice; bottom - HGF<sup>ex.5 flox</sup>;Mx1-cre mice. (<b>C</b>) RT-PCR shows the presence of both recombined and unrecombined HGF mRNA in KO livers. (<b>D</b>) WB for HGF in control and HGF KO livers shows no differences after recombination. Ponceau represents loading control.</p
Suppression of spontaneous proliferation of mouse hepatocytes in the presence of 10
<p><sup>−<b>6</b></sup><b> M dexamethasone.</b> Tritiated thymidine was added continually in the cultures, from 4 hours after cell plating following perfusion of the liver by collagenase. Cultures were harvested at the indicated time points in the X-axis. Addition of HGF and EGF is indicated as “GFs”. (For growth factor concentration, please see Materials and Methods).</p
List of putative SUMO2 substrate proteins that are reduced in abundance in the purified infected compared to uninfected samples.
<p>The entries are shaded by degree of change, and all gave H/L ratios of 2 or greater with SigB values <0.1. The darkest shading indicates greater than 10-fold decrease, then in order of shading 7–10 fold, 5–7 fold, 3–5 fold and 2–3 fold.</p
Core PML-NB proteins, but not IFI16, associate with infecting HSV-1 genomes.
<p>HFt cells were mock or infected with either HSV-1<sup>EdU</sup> or HSV-1<sup>EdC</sup> at an MOI of 3 PFU/cell. Cells were fixed and permeabilized at the indicated times (mpi; post-addition of virus). vDNA and PML-NB host factors were detected by click chemistry and indirect immunofluorescence staining, respectively. (A-E) Localization of PML (green), and either Daxx, Sp100, ATRX, SUMO2/3 (SUMO), or IFI16 (cyan; as indicated) to infecting HSV-1<sup>EdU</sup> vDNA (red, white arrows). Insets show magnified regions of interest (dashed boxes) highlighting host protein localization with vDNA. Cut mask (yellow) highlights regions of colocalization between host proteins and vDNA (as indicated). Weighted colocalization coefficients are shown. Individual channel images shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1006769#ppat.1006769.s003" target="_blank">S3 Fig</a>. Nuclei were stained with DAPI (blue). (F) Quantitation of host protein recruitment to infecting viral genomes labelled with EdU (as shown in A-E) or EdC. Boxes: 25<sup>th</sup> to 75<sup>th</sup> percentile range; black line: median weighted (w.) colocalization coefficient; whiskers: 5<sup>th</sup> to 95<sup>th</sup> percentile range. Solid line indicates coincident threshold level (weighted colocalization coefficients < 0.2). n ≥ 50 vDNA foci per sample population from a minimum of three independent infections.</p
Degradation of selected cellular proteins by ICP0 in the absence of infection.
<p>A. HA-TetR and HA-cICP0 cells were treated with doxycycline (100 ng/ml) for 24 h (+) or left untreated (-), then whole cell extracts were prepared and analyzed by western blotting for the indicated proteins using antibodies directed against the endogenous proteins. The dashes on the right indicate the major presumed specific bands (determined on the basis of analyses in previous figures) in each case. Except in the cases of CITED2 and ZBTB4, these major bands decrease in the presence of ICP0. The upper dash in the ZBTB4 panel indicates a potential sumoylated form that is sensitive to ICP0. (B). Analysis of HA-cICP0 cells (left, lanes 1 and 2) and derivatives of HA-cICP0 cells that had been transduced with lentivirus vectors expressing myc-tagged BEND3 (left, lanes 3 and 4) and MBD1 (right), before and after induction of ICP0 expression, illustrating that ICP0 is sufficient to degrade BEND3 and MBD1.</p
List of putative SUMO2 substrate proteins that are reduced in abundance in the purified infected compared to uninfected samples.
<p>The entries are shaded by degree of change, and all gave H/L ratios of 2 or greater with SigB values <0.1. The darkest shading indicates greater than 10-fold decrease, then in order of shading 7–10 fold, 5–7 fold, 3–5 fold and 2–3 fold.</p