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Consistent with robust p38 activation during ischaemia, we saw strong activation of both MKK3 and MKK6.
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Similarly to the perfused heart experiments, we saw a strong activation of ASK1 during ischaemia which correlated with increased p38 activity (Fig. 5B).
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Altogether, these results support the relevance of sCD1d-antitumor fusion proteins for cancer therapy, as seen by the strong activation of human iNKT cells, revealed by direct tumor cytotoxicity and cytokine release.
This was in contrast to the strong activation seen with the positive control ALK-F1174L whish is a well characterized constitutively active ALK neuroblastoma mutation.
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However, under the short-time course examined here, in which a clear autophagic response was seen, we did not note strong activation of p53 as measured by increase in p53 levels, or enhanced expression of another p53-target gene, p21 (data not shown).
The notion that there is some background staining in unstimulated 293T cells, was reported previously (Staal et al, EMBO REP) and with strong activation signals a clear increase is seen.
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