Low iron activates Furin which sheds to cleave the prodomain of ADAM10 rendering it activated.
<p><b>(A.i.)</b> Western blot of whole cell lysate of uninfected-MФs, LD-R, and LD-S infected-MФs at 24 hrs pi showing the expression of precursor-ADAM10 (p-ADAM10, ~90 kDa) and mature-ADAM10 (m-ADAM10, ~64 kDa) keeping β-actin acts as a housekeeping control. <b>(A.ii.)</b...
محفوظ في:
| المؤلف الرئيسي: | |
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| مؤلفون آخرون: | , , , |
| منشور في: |
2025
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| الموضوعات: | |
| الوسوم: |
إضافة وسم
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| الملخص: | <p><b>(A.i.)</b> Western blot of whole cell lysate of uninfected-MФs, LD-R, and LD-S infected-MФs at 24 hrs pi showing the expression of precursor-ADAM10 (p-ADAM10, ~90 kDa) and mature-ADAM10 (m-ADAM10, ~64 kDa) keeping β-actin acts as a housekeeping control. <b>(A.ii.)</b> Bar graph representing the relative intensity of mature-ADAM10 (m-ADAM10) vs precursor-ADAM10 (p-ADAM10). Significant upregulation of relative intensity of m-ADAM10 with respect to p-ADAM10 in LD-R 24 hrs pi as compared to both UI and LD-S 24hrs pi (****, <b>P</b> ≤ 0.0001) whereas no change is observed between LD-R 24 hrs pi and UI (ns, <b>P</b> > 0.05). <b>(B.i.a.)</b> Western blot of whole cell lysate showing furin expression of DFO-treated-MФs, LD-S, and LD-R infected-MФs at 24 hrs pi. <b>(B.i.b.)</b> Bar graph representing the relative intensity of Furin, i.e., fold change with respect to β-actin for whole cell lysate. Significant enrichment of Furin observed in LD-S as compared to both DFO-treated control and LD-R 24 hrs pi (****, <b>P</b> ≤ 0.0001) whereas no change is observed between LD-R 24 hrs pi and DFO-treated control (ns, <b>P</b> > 0.05). Each densitometry analysis is presented as a bar graph of Mean ± SEM for 3 biological replicates. <b>(B.ii.)</b> 40X Confocal images showing the localization of furin (red) in uninfected-MФs, LD-S, and LD-R infected-MФs. <b>(B.iii.)</b> Western blot of furin expression in the supernatant fraction of same experiments sets as B.i. <b>(C.)</b> Confocal images showing Furin (red, 3<sup>rd</sup> panel and edges, 6<sup>th</sup> panel), ADAM10 (purple), and CD11b (green) colocalization in LD-S and LD-R infected-MФs at 24 hrs pi. One representative small nucleus of LD has been marked in (<b>*</b>) to show the infected-MФs. Scale bars indicate 20µm. <b>(D.)</b> Scheme showing Furin maturation starting from Endoplasmic reticulum (ER) where it remains inactivated (observed in LD-S-infected-MФs at 24 hrs pi) to getting shed as an active enzyme (observed in LD-R-infected-MФs at 24 hrs pi) to cleave prodomain of p-ADAM10 rendering it activated (m-ADAM10), which in turn cleaves extracellular domain of SIRPα in LD-R-infected-MФs at 24 hrs pi.</p> |
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