Hello! Thank you for your question. At this time I do not have an exact number for you. We have stained for macrophages and smooth muscles within the plaques as shown in figure 8. We did not see any difference in plaque macrophage staining intensity between wildtype or lipin-1mKO. I have not measured dendritic cells or neutrophils yet; but I have isolated and digested aortas from WT and lipin-1mKO mice and ran flow cytometry to look at cell numbers of macrophages, neutrophils, dendritic cells, t cells, and b cells. I have not yet analyzed the data. I would hypothesize that macrophages would have been the most prevalent but I did not see that in my staining. Therefore it might not be a difference in the cell numbers but rather how the macrophages are responding in the plaque.
Have there been studies to show (or have you looked yourself) to see if Cre expression alone in macrophages, dendritic cells, and neutrophils alters vascular biology?
Hi Sandeep! Thanks for the question. We have not looked directly to see if Cre expression alone alters vascular biology and I cannot recall any studies that have shown that. I would not hypothesize that it would not cause any drastic affects because the cre should only excise at loxP sites. I will mention that with our studies we make sure to use littermate controls. Therefore, our wildtype mice will still have the lipin-1 gene flanked by the lox site (they just won’t have cre expression).
Do you know the cell number of macrophages, dendritic cells and neutrophils in plaques? which is the most and which is the least? Thanks.
Hello! Thank you for your question. At this time I do not have an exact number for you. We have stained for macrophages and smooth muscles within the plaques as shown in figure 8. We did not see any difference in plaque macrophage staining intensity between wildtype or lipin-1mKO. I have not measured dendritic cells or neutrophils yet; but I have isolated and digested aortas from WT and lipin-1mKO mice and ran flow cytometry to look at cell numbers of macrophages, neutrophils, dendritic cells, t cells, and b cells. I have not yet analyzed the data. I would hypothesize that macrophages would have been the most prevalent but I did not see that in my staining. Therefore it might not be a difference in the cell numbers but rather how the macrophages are responding in the plaque.
Have there been studies to show (or have you looked yourself) to see if Cre expression alone in macrophages, dendritic cells, and neutrophils alters vascular biology?
Hi Sandeep! Thanks for the question. We have not looked directly to see if Cre expression alone alters vascular biology and I cannot recall any studies that have shown that. I would not hypothesize that it would not cause any drastic affects because the cre should only excise at loxP sites. I will mention that with our studies we make sure to use littermate controls. Therefore, our wildtype mice will still have the lipin-1 gene flanked by the lox site (they just won’t have cre expression).