Reactive Oxygen species scavenging activity of PRME and the reference compounds.

<p>(A) Hydroxyl radical inhibition, (B) hypochlorous radical scavenging, (C) superoxide radical inhibition, (D) singlet oxygen radical scavenging. The results are mean ± S.D. of six parallel measurements. **p<0.01 and ***p<0.001 vs. 0 µg/ml.</p

Trolox equivalent antioxidant capacity and IC₅₀ values of the lichen extract (PRME) and standard compounds for ROS and RNS scavenging.

<p>₅₀ values of all activities are determined in µg/ml. Data expressed as mean ± S.D (n = 6). EDTA represents Ethylenediamine tetraacetic acid.^# IC</p><p> p<0.001.</p

Proposed mechanisms of PRME-induced S and G2/M phase cell cycle arrest and apoptosis in MCF-7 cells.

<p>Proposed mechanisms of PRME-induced S and G2/M phase cell cycle arrest and apoptosis in MCF-7 cells.</p

HPLC chromatogram of PRME.

<p>Inset shows expanded region of the chromatogram with retention time of 4.5–10 minutes. Peaks marked signify the retention peak of purpurin (2.4 min), catechin (3.13 min), tannic acid (3.64 min) and reserpine (4.68 min).</p

Qualitative and quantitative phytochemical analysis of PRME.

<p>“+” represents presence of the phytoconstituent; “−” represents absence of the phytoconstituent; “ND” represents “Not determined”. Phen- Phenol, Flav- Flavonoid, Carbo- Carbohydrate, Tan.- Tannin, Alka- Alkaloid, Asc- Ascorbic acid, Ter- Terpenoids, Triter- Triterpenoids, Anth-Anthraquinones, Sap- Saponin, Gly- Glycoside; Total phenolics (mg/100 mg extract gallic acid equivalent), Total flavonoids (mg/100 mg extract quercetin equivalent), Carbohydtrate (mg/100 mg extract glucose equivalent), Tannin (mg/100 mg extract catechin equivalent). Alkaloid (mg/100 mg extract reserpine equivalent), Ascorbic acid (mg/100 mg extract L-ascorbic acid equivalent) </p

Flow cytometric cell cycle distribution of PRME treated MCF-7 cells with increasing time.

<p>Sub-G1, G1, S, and G2/M phases of PRME (300 µg/ml) treated MCF-7 cells at (A) 0 hour, (B) 6 hours, (C) 12 hours, (D) 24 hours, (E) 36 hours, (F) 48 hours. (G) Graphical representation of % cell population in different phases.</p

Western blot analysis of cell cycle related proteins of MCF-7 cells treated with 300 µg/ml PRME.

<p>Graphs adjoining the blots represent the expression levels of corresponding proteins for indicated time intervals: (A) Cyclin B1, (B) Cdk-2, (C) Cdc25c, (D) Cdk-1, (E) Cyclin A1, (F) p53, (G) p21.</p

Western blot analysis of apoptosis related proteins of MCF-7 cells treated with 300 µg/ml PRME.

<p>Graphs adjoining the blots represent the expression levels of corresponding proteins for indicated time intervals: (A) Pro and cleaved caspase-9, (B) Pro and cleaved caspase-3, (C) Native and cleaved PARP, (D) Bax and Bcl-2, (E) Pro and cleaved caspase-8, (F) Bid and t-Bid.</p

Flow cytometric plots of Annexin-V-FLUOS and PI staining of PRME treated MCF-7 cells with increasing doses.

<p>MCF-7 cells were treated for 48 hours with different concentrations: (A) Control, (B) 50 µg/ml, (C) 80 µg/ml, (D) 100 µg/ml, (E) 200 µg/ml, (F) 300 µg/ml of PRME. Numbers in boxes represent % of total cells in respective boxes.</p

DPPH radical scavenging activity of PRME and the reference compound.

<p>The results are mean ± S.D. of six parallel measurements. **p<0.01 and ***p<0.001 vs. 0 µg/ml.</p