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Thus, modulation of hepcidin expression by endogenous BMPs does not seem to underlie the reduction of hepcidin mRNA by DMOG and DP.
To determine whether either of these HIF-α subunits is necessary for the modulation of hepcidin mRNA levels we transfected HepG2 cells with siRNAs directed against HIF-1α or HIF-2α.
Since this pattern of TfR2 regulation matched the modulation of hepcidin transcripts, in particular the reduction by DMOG and DP, regulation of TfR2 expression may contribute to the observed responses of hepcidin expression in vitro.
As a first step to delineate the molecular mechanism(s) involved in the UPR-dependent modulation of hepcidin expression, the levels of this transcription factor were evaluated during the time- and dose-response assays.
It should also be noted that the profound biomedical implications associated with the modulation of hepcidin pathways render the hepcidin ferroportin axis an attractive target for drug development.
These preclinical data suggest that modulation of hepcidin expression could be a substitute for or an adjunct to iron chelation therapy in patients with thalassemia.
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In terms of kinetics, the down-modulation of hepcidin initiated after 5 h of treatment (Fig. 2B) also overlapped with both induction of CHOP (Fig. 1D) and reduction of C/EBPα (Fig. 4A) proteins.
This suggests that CHOP may not be involved in the down-modulation of hepcidin by EPO in physiological conditions.
In conclusion, the present results support the hypothesis of a direct role for EPO in the regulation of hepcidin, via modulation of C/EBPα expression.
Although the kinetics of C/EBPα binding did not strictly match the expression profile of hepcidin, our fEMSA results suggest that modulation of C/EBPα binding to hepcidin promoter might underlie the up-regulation of its expression detected after 24 h of DTT treatment.
The central role of hepcidin in iron homeostasis, coupled to its marked modulation in HepG2 cells undergoing ER stress, prompted us to focus on this molecule.
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