The dose dependent cytotoxic effect of 1j bis-lawsone derivative and CDDP on CCF-4 and NKE cells.

<p>a. Effects of different concentrations of <b>1j</b> ranging from 0 μM to 50μM, on both the cells viability by MTT assay. b. Effects of different concentrations of CDDP ranging from 0 μM to 50μM, on both the cells viability by MTT assay. c. Cytotoxic effect of both <b>1j</b> and CDDP on both the cancerous and normal cell line by LDH leakage assay. Each point represents mean ±SD, n = 3 (number of plates). ‘‘*” represents the significant difference compared to the control cells. (P* < 0.05).</p

IC₅₀ of the synthesized 1j derivative on six different cell lines.

<p>IC₅₀ of the synthesized 1j derivative on six different cell lines.</p

Effect of 20 μM 1j derivative on cell migration was observed by a phase contrast microscope (100X).

<p>a. untreated CCF-4 cells b. 1j exposed CCF-4 cells. Data are representative of three independent experiments. All data are mean ± SD, for 3 independent experiments and were analyzed by one-way ANOVA. ‘‘*” represents the significant difference between the normal control and <b>1j</b> exposed cells.</p

Selective Pro-Apoptotic Activity of Novel 3,3′-(Aryl/Alkyl-Methylene)Bis(2-Hydroxynaphthalene-1,4-Dione) Derivatives on Human Cancer Cells via the Induction Reactive Oxygen Species - Fig 14

<p>a. Immunoblot analysis of different signalling molecules considered in the study. β actin was used as an internal control. Data are representative of three independent experiments. Control: untreated cells, 1j: 20 μM <b>1j</b> exposed cells. b. Densitometric analysis of the respective immunoblots. All data are mean ± SD, for 3 independent experiments and were analyzed by one-way ANOVA. ‘‘*” represents the significant difference between the normal control and <b>1j</b> exposed cells. c. Schematic representation of the most probable signalling cascade moduated due to <b>1j</b> exposure.</p

Effect of 20 μM 1j and 25 μM CDDP on cell cycle progression in CCF-4 cells.

<p>It was observed that <b>1j</b> derivative and CDDP induces cell cycle arrest at G0-G1 phase. Data are representative of three independent experiments. All data are mean ± SD, for 3 independent experiments and were analyzed by one-way ANOVA. ‘‘*” represents the significant difference between the normal control and <b>1j</b> exposed cells.</p

Effect of 20 μM 1j and 25 μM CDDP on ROS production in the cancerous and normal cells.

<p>a-b. DCFH-DA staining shows increased ROS production upon exposure of both the compounds in CCF-4 and NKE cells. c-d. Differential induction of ROS on CCF-4 and NKE cells upon exposure to 20 μM <b>1j</b> and 5mM NAC. e. Intracellular ROS production was detected by changes in the fluorescence intensity of DCF by fluoroscent microscopy (20X). Data are representative of three independent experiments. f. Effect on cell viability of CCF-4 cells upon exposure to 20 μM <b>1j</b> and 5mM NAC. Each column represents mean ±SD, n = 6. ‘‘*” represents the significant difference between the normal control and <b>1j</b> treated cells (P* < 0.05).</p

Chemical structures of the synthesized bis-lawsone compounds.

<p>Chemical structures of the synthesized bis-lawsone compounds.</p

The dose dependent effect on cell viability of all the 22 bis-lawsone derivatives on the 5 human cancer cell lines.

<p>The derivatives were exposed to the different cells at a dose of 5, 10, 20 & 40 μM. a. CCF-4 cells, b. A549 cells, c. HeLa cells, d. SKRC-45 cells, e. A498 cells. Each point represents mean ±SD, n = 3 (number of plates).</p

Detection of 1j and CDDP induced apoptosis on both the CCF-4 and NKE cells via Annexin V-FITC staining.

<p>The cells were incubated separately with desired molecules (20 μM <b>1j</b> and 25 μM CDDP) followed by staining with Annexin V-FITC and then the cells were flow cytometrically analyzed. Dual parameter dot plot of FITC-labelled Annexin V fluorescence (x-axis) has been shown in logarithmic fluorescence intensity. Data are representative of three independent experiments.</p

Effect of 20 μM 1j and 25 μM CDDP on mitochondrial membrane potential in CCF-4 cells and NKE cells as shown by the monomeric green fluorescence of the JC-1 dye.

<p>Data are representative of three independent experiments.</p