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nnm-2021-0161 - Supplementary Figures S1-S14.pdf

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posted on 16.08.2021, 13:38 by Pragya Prasanna, Prakash Kumar, Saptarshi Mandal, Tanvi Payal, Saurabh Kumar, Ugir Hossain Sk., Prolay Das, Ravichandiran V, Debabrata Mandal
Supplementary Figure S1. A scheme for the synthesis of DHF-GNP using DHF and HauCl4 solution.

Supplementary Figure S2. Characterization of DHF-GNPs by Zeta potential studies (A) and by DLS studies (B).

Supplementary Figure S3. Characterization of c-GNPs by DLS studies and (A) Transmission electron microscopy (B).

Supplementary Figure S4. The amount of DHF released from 10 mM DHF-GNP was measured for 72 h after incubation in different pH of PBS (A) and after incubation in PBS (pH 6.0) and blood plasma (pH 7.4) (B).

Supplementary Figure S5. Dose-response curve of DHF, DHF-GNP, and c-GNP after 72 h treatment against promastigotes in vitro(A) and intracellular amastigotes ex vivo (B).

Supplementary Figure S6. Stability studies using a dose-response curve of 6 months old DHF, DHF-GNP, and c-GNP after 48 h treatment against promastigotes in vitro (A) and amastigotes ex vivo (B).

Supplementary Figure S7. Hemolysis assay on human red blood cells after 4 h treatment with

DHF-GNP with AmB and 0.1% DMSO as positive and negative control respectively.

Supplementary Figure S8. Semiquantitative PCR for different genes on promastigotes after treatment with DHF and DHF-GNP at IC50 dose for 24 h (A) and after treatment with 2 × IC50 doses for 8 h (B).

Supplementary Figure S9. Semiquantitative PCR for genes of amastigote-infected macrophages after treatment with DHF and DHF-GNP with 2 × IC50 doses for 8h.

Supplementary Figure S10. Effect of different doses of DHF and DHF-GNP on the arginase activity in treated promastigote cell lysates.

Supplementary Figure S11. Uptake of gold nanoparticles by promastigotes as detected by AAS.

Supplementary Figure S12. Enzyme assay with LD promastigotes lysates after treatment with DHF (120-240 μM), DHF-GNP (40-80 μM), and c-GNP (20μM). Superoxide dismutase assay and (A) Ascorbate peroxidase assay (B).

Supplementary Figure S13. Antioxidant activities of DHF (5-80 μM), DHF-GNPs (5-80 μM) measured by percentage inhibition of DPPH scavenging activity(A) Determination of reactive oxygen species in LD promastigotes treated with DHF (40-280 μM), DHF-GNP (40-280 μM), and c-GNP (5 μM) for 12 hours (B).

Supplementary Figure S14. Estimation of reactive nitrogen species by Griess reagent after DHF, DHF-GNP, and c-GNP treatment of macrophages. Cells are pre-treated with DPI (100 µM) whenever required.

Funding

The funding for this research work was provided by the Ministry of Chemicals and Fertilizers, Government of India.

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