Sentences with phrase «ko cells»

Roman numerals represent different microscopic slides and Arabic numerals different cell lines labelled as follows: (I) wt cells (1) labelled with 2 - μm beads, XpdTTD / TTD cells (2) with 0.79 - μm beads, and XpdTTD / KO cells (3) with no beads; (II) wt cells (1) labelled with 0.79 - μm beads and XpdTTD / † XPCS cells (4) with no beads; and (III) wt cells (1) labelled with 0.79 - μm beads and XpdTTD / † XP cells (5) with no beads.

In accordance with the gene dosage, a further reduction of up to 70 % of the wt level was observed in hemizygous XpdTTD / KO cells.

Significant effects of compound heterozygosity on NER subpathways relative to XpdTTD / KO cells were observed in XpdTTD / † XP cells but only for UV - UDS activity.

Capsaicin induced a further induction of TPA - increased COX - 2 expression in EGFR / WT cells, but not in EGFR / KO cells.

EGFR / WT and KO cells were cultured overnight and starved for 24 h. Cells were treated with capsaicin (50 μmol / L) for 30 min and then TPA (20 ng / mL) for 2 h. Cells were lysed and cox - 2 mRNA levels were analyzed by RT - PCR.

Each lysate product is sold in kit format consisting of a KO cell lysate and a parental cell lysate, which are immediately available from OriGene for the academic and industrial research markets.

Not exact matches

EGFR / WT and KO MEFs were treated with capsaicin (0, 10, or 50 μmol / L) for 30 min followed by TPA (20 ng / mL) for 4 h. Cells were lysed and COX - 2 expression analyzed by Western blot.

For instance, ZBTB48 KO (knock - out) cells loose the expression of MTFP1 leading to defects in the mitochondrial network with mitochondria clustering around the nucleus instead of being widely spread throughout the cell.

Derived mostly from human embryonic kidney 293T (HEK293T) and HeLa cell lines, EdiGene Knockout (KO) Cell Lysates have been optimized through the use of genome editing technology and validated at the genomic level through PCR and Sanger - sequencing techniques to ensure the accuracy and knockout of the target gcell lines, EdiGene Knockout (KO) Cell Lysates have been optimized through the use of genome editing technology and validated at the genomic level through PCR and Sanger - sequencing techniques to ensure the accuracy and knockout of the target gCell Lysates have been optimized through the use of genome editing technology and validated at the genomic level through PCR and Sanger - sequencing techniques to ensure the accuracy and knockout of the target gene.

First, we produced PARP1 - KO ovarian cancer cell lines using CRISPR / Cas9 gene editing to test the loss of PARP - 1 as a resistance mechanism to all clinically used PARP inhibitors.

If interactions between IL - 15 and IL - 15Rα were to occur strictly as a secreted cytokine binding to a cell surface receptor, then similar exercise and muscle phenotypes would have been observed in both IL - 15Rα — KO mice and IL - 15 — KO mice, since ligand - receptor binding would have been interrupted in both of these mouse strains.

Nonetheless, responses in KO animals to most stimuli (100 mm NH4Cl, 500 mm sucrose, 300 mm MSG with amiloride, 5 — 20 mm citric acid, and 5 — 20 mm HCl) were reduced compared with WT (Fig. 6B, Table 4), suggesting a role for 5 - HT3 signaling in more than just acid and salt transmission; that is, tastants likely to evoke direct type III cell - mediated signaling.

However, closer scrutiny of these studies raises questions about the role of CD4 + T cells as the sole mediators of corneal graft rejection, as corneal allografts undergo immune rejection in 33 % of the mice and 64 % of the rats treated with anti-CD4 monoclonal antibody (12, 13) and in 45 % of the CD4 KO mice (14).

The capacity of CD8 − T cells from CD4 KO donors to mediate corneal allograft rejection is puzzling and on the surface, counterintuitive, since these cells are presumably double negative (DN) T cells.

Spleen cell suspensions were collected 7 — 14 days after CD4 KO mice rejected BALB / c corneal allografts.

Number of experiments: n = 15 (XpdTTD / TTD), n = 6 (XpdTTD / KO), n = 4 (XpdTTD / † XPCS), n = 2 (XpdTTD / † XP); 1 — 2 cell lines per genotype were included in each experiment.

By contrast, 71 % (5/7) of the beige nude mice that received spleen cells from CD4 KO mice rejected their BALB / c corneal allografts.

The present demonstration of T cell - mediated apoptosis of allogeneic corneal cells from CD4 KO mice is consistent with previous findings, which noted the presence of apoptotic keratocytes and corneal endothelial cells in rejected corneal allografts in humans and rats respectively (5, 32).

In our study, CD8 − T cells from CD4 KO rejector mice failed to display CTL or DTH activity, yet they were capable of inducing donor - specific apoptosis of corneal endothelial cells.

Spleen cells were collected from CD4 KO mice 7 — 14 days after they had rejected BALB / c corneal allografts.

The results of a typical experiment are shown in Figure 3 and demonstrate that even though spleen cells from CD4 KO mice could adoptively transfer corneal allograft rejection, they did not display conventional CTL activity against either BALB / c corneal epithelial or endothelial cells.

The present findings are derived from studies using CD4KO mice and thus, raise the question as to whether the CD4 + T cell - independent immune mechanisms in CD4 KO mice differ from those involved in corneal allograft rejection in wild - type mice whose CD4 + T cells population have been depleted with monoclonal antibodies.

The role of DN T cells in corneal allograft rejection was confirmed in two separate in vitro assays in which CD8 − cells were isolated from CD4 KO donors that had rejected corneal allografts and were found to induce apoptosis of donor - specific corneal cells.

The CD8 + T cells were already primed, as they were collected from CD4 KO mice that had rejected corneal allografts and thus, may have functioned differently from CD8 + T cells from naïve CD4 KO mice.

Indeed, our results indicate that adoptive transfer of CD8 + T cells from CD4 KO mice that had rejected corneal allografts resulted in the rejection of 95 % of the corneal allografts transplanted to athymic recipients.

In CD4 KO mice, T cell - independent rejection can involve either CD8 + or CD8 − T cells.

Histopathological examination of rejected corneal allografts in CD4 KO mice revealed a mixed inflammatory infiltrate containing large numbers of neutrophils and mononuclear cells, which was indistinguishable from the infiltrate seen in rejected corneal allografts in wild - type mice (data not shown).

Relative to XpdTTD / KO hemizygote cells, UV survival was improved by the homozygous lethal Xpd † XPCS allele in XpdTTD / † XPCS compound heterozygous cells and to a lesser degree by the Xpd † XP allele (Figure 4A).

However, in vitro assays using spleen cells from CD4 KO mice that had rejected BALB / c corneal allografts failed to detect CTL activity against donor corneal epithelial or endothelial cells.

The extensive apoptosis of allogeneic corneal endothelial cells by CD8 − T cells from CD4 KO mice was perplexing, as this cell population should not contain CD4 + cells.

Briefly, single - cell suspensions of C57BL / 6 KO and wild - type lymphocytes, which had undergone a 96 hr in vitro boost with γ - irradiated (3,000 rad) BALB / c stimulator lymphocytes, were added to 96 - well microtiter plates (Corning Inc., Corning, NY) along with 2 × 104 51Cr - labeled BALB / c corneal epithelial or endothelial cells in a total volume of 200 μl / well.

(E) Xpd dose - dependent reduction of TFIIH in homozygous XpdTTD / TTD, hemizygous XpdTTD / KO, and compound heterozygous XpdTTD / † XPCS and XpdTTD / † XP cells by comparative immunofluorescence of the p62 subunit of TFIIH.

Both CD8 − and CD8 + T cells from CD4 KO corneal allograft rejector mice mediated corneal allograft rejection following adoptive transfer to nude mice.

BALB / c corneal allografts were transplanted to C57BL / 6 beige nude mice that received either CD8 − or CD8 + T cells from C57BL / 6 CD4 knockout (KO) mice that had rejected BALB / c corneal allografts.

Spleen cells were isolated 7 — 14 days after C57BL / 6 CD4 KO mice had rejected BALB / c corneal allografts and were tested for anti - BALB / c CTL in a conventional 4 - hr 51Cr - release assay using BALB / c corneal epithelial and endothelial target cells.

Moreover, we found that T cells from double - knockout (DKO) mice did not demonstrate enhanced tumor activity above that observed in DGKζ single - knockout (KO) mice.

Together, these data indicate that peripheral T cell numbers are relatively preserved in KO or DKO mice compared with WT mice but that fewer naive CD8 + T cells are present in all genotypes when compared with WT.

Additionally, we identified that combined deletion of DGKζ and Cbl - b resulted in similar impairment of antigen - specific memory CD8 + T cell generation and / or maintenance compared with KO.

Importantly, evaluation of activation markers on peripheral CD8 + T cells showed decreased numbers of naive (CD44loCD62Lhi) T cells in splenocytes of KO mice (Fig. 2C), as has been previously observed (17), which was enhanced in DKO mice.

We next evaluated T cell numbers in peripheral immune organs and found comparable numbers of CD8 + and CD4 + T cells in spleens of WT mice compared with KO or DKO mice with some modest differences in CD8 + and CD4 + T cell percentages between genotypes (Fig. 2B).

In case of the Sanger Institute, lines of interest can be nominated from the Sanger ES cell collection from which then a KO mouse line will be produced and phenotyped.

On the other hand, the SCP1 expression is detected in germ cells of SCP3 KO mice, though the structure of the synaptonemal complex and localization of SCP1 in the complex are abnormal in comparison to the wild - type germ cells [50].

In Thy1 - PE labeling experiments, Ccr6 KO mice showed reduced frequencies of labeled cells, a result that was most significant for the Vγ4 + population (Figure 4G).