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Treatment of macrophages with apoptotic cells or cell supernatant
Last updated date: Jun 30, 2020 Views: 1096 Forks: 0
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Dear Parul,
In page 3, the second paragraph, we say
"Treatment of the apoptotic cell supernatant with proteinase K (50 μg/ml for 60 min), DNase I (6 U/ml
for 60 min), or RNase A (5 μg/ml for 60 min) did not prevent its ability to enhance Thbs1 gene expression(Figure 1D),"
No effect on the gene expression in macrophages indicates that these enzymes have not damaged the macrophage function.
(We have not removed these enzymes before adding to macrophages).
This anser to you qestions 1 and 2.
For question 3, please see the Figure 1 legend.
Figure 1. Factor(s) released from apoptotic cells stimulate gene expression in macrophages. (A and B) BMDMs
were incubated for 1 hr with medium or with the supernatant of W3 cells that had been treated with (apoptotic) or
without (living) 30 units/ml FasL. RNA from BMDMs was then subjected to microarray analysis
Sincerely Yours,
Shigekazu Nagata
- Nagata, S(2020). Treatment of macrophages with apoptotic cells or cell supernatant. Bio-protocol Preprint. bio-protocol.org/prep348.
- Yamaguchi, H., Maruyama, T., Urade, Y. and Nagata, S.(2014). Immunosuppression via adenosine receptor activation by adenosine monophosphate released from apoptotic cells. eLife. DOI: 10.7554/eLife.02172
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