Journal: International Journal of Molecular Sciences

Article Title: Targeting Voltage-Gated Potassium Channels in Breast Cancer: Mechanistic Insights into 4-Aminopyridine-Induced Cell Death

doi: 10.3390/ijms26167768

Figure Lengend Snippet: Determination of the IC 50 values of 4-AP in L929 and MCF-7 cell lines. Concentrations of 25 mM, 12.5 mM, and 6.25 mM were used as the highest doses. Since the expected IC 50 range was between 1 and 5 mM, finer dose increments of 1 mM were applied within this range. Cell viability was assessed using the trypan blue exclusion method with a hemocytometer. Following viability measurements, inhibitor vs. response curves were plotted to calculate IC 50 values. The IC 50 value of 4-AP was determined to be 5 mM in L929 cells ( a ) and 4 mM in MCF-7 cells ( b ). Data represent mean ± SEM from more than three independent experiments ( n ≥ 3).

Article Snippet: The breast cancer cell line MCF-7 (ATCC HTB 22) and healthy mouse fibroblast cell line L929 (ATCC CCL-1) were used in this study.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Targeting Voltage-Gated Potassium Channels in Breast Cancer: Mechanistic Insights into 4-Aminopyridine-Induced Cell Death

doi: 10.3390/ijms26167768

Figure Lengend Snippet: Assessment of cell viability following CHX and 4-AP treatments. Cells were treated with the IC 50 concentration of 4-AP, and cell viability was assessed using the trypan blue exclusion method with a hemocytometer. For CHX treatment, a 20 µM dose was selected. Cells were pre-incubated with CHX for 1 h prior to 4-AP exposure. CHX pre-treatment resulted in increased cell viability in both the L929 ( A ) and MCF-7 ( B ) cell lines. *** p < 0.001, and **** p < 0.0001, n ≥ 3.

Article Snippet: The breast cancer cell line MCF-7 (ATCC HTB 22) and healthy mouse fibroblast cell line L929 (ATCC CCL-1) were used in this study.

Techniques: Concentration Assay, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Targeting Voltage-Gated Potassium Channels in Breast Cancer: Mechanistic Insights into 4-Aminopyridine-Induced Cell Death

doi: 10.3390/ijms26167768

Figure Lengend Snippet: Assessment of cell viability following Z-VAD-FMK and 4-AP treatments. Cells were treated with 50 µM Z-VAD-FMK, either alone or in combination with 4-AP. Cell viability was assessed using the trypan blue exclusion method with a hemocytometer. Treatment with Z-VAD-FMK alone resulted in a slight decrease in cell viability compared to the control group. However, pre-treatment with Z-VAD-FMK for 1 h prior to 4-AP exposure significantly reduced cell viability in both L929 ( A ) and MCF-7 ( B ) cell lines. ** p < 0.01, **** p < 0.0001, n ≥ 3.

Article Snippet: The breast cancer cell line MCF-7 (ATCC HTB 22) and healthy mouse fibroblast cell line L929 (ATCC CCL-1) were used in this study.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Targeting Voltage-Gated Potassium Channels in Breast Cancer: Mechanistic Insights into 4-Aminopyridine-Induced Cell Death

doi: 10.3390/ijms26167768

Figure Lengend Snippet: Assessment of cell viability following 2-APB and 4-AP treatments. Cells were treated with 10 µM 2-APB, either alone or in combination with 4-AP. Cell viability was assessed using the trypan blue exclusion method with a hemocytometer. Pre-treatment with 2-APB for 1 h increased cell viability in both L929 ( A ) and MCF-7 ( B ) cell lines. *** p < 0.001, and **** p < 0.0001, n ≥ 3.

Article Snippet: The breast cancer cell line MCF-7 (ATCC HTB 22) and healthy mouse fibroblast cell line L929 (ATCC CCL-1) were used in this study.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Targeting Voltage-Gated Potassium Channels in Breast Cancer: Mechanistic Insights into 4-Aminopyridine-Induced Cell Death

doi: 10.3390/ijms26167768

Figure Lengend Snippet: Percentage changes in intracellular Ca 2+ levels following treatments. Intracellular calcium concentrations were measured using the Fura-2 fluorescent dye. Percentage changes in Ca 2+ levels were calculated relative to the untreated control group. Treatment with 4-AP and its combinations increased intracellular Ca 2+ levels. CHX and Z-VAD-FMK alone did not induce significant increases, whereas 2-APB alone elevated intracellular Ca 2+ concentrations in both L929 ( A ) and MCF-7 ( B ) cell lines. * p < 0.05, *** p < 0.001, and **** p < 0.0001, n ≥ 3. Non-significant results are not indicated in the figure.

Article Snippet: The breast cancer cell line MCF-7 (ATCC HTB 22) and healthy mouse fibroblast cell line L929 (ATCC CCL-1) were used in this study.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Targeting Voltage-Gated Potassium Channels in Breast Cancer: Mechanistic Insights into 4-Aminopyridine-Induced Cell Death

doi: 10.3390/ijms26167768

Figure Lengend Snippet: Effect of drug treatments on membrane polarization. To assess the effects of elevated intracellular Ca 2+ levels on membrane potential, DiBAC 4 (3) fluorescent dye was used. Since CHX and Z-VAD-FMK did not induce significant increases in intracellular Ca 2+ levels, they were excluded from DiBAC 4 (3) measurements. Treatment with 4-AP and its combinations increased membrane depolarization, with the most pronounced effect observed following co-treatment with 2-APB. In contrast, 2-APB alone did not cause substantial depolarization in either L929 ( A ) or MCF-7 ( B ) cell lines. **** p < 0.0001, n ≥ 3. Non-significant results are not indicated in the figure.

Article Snippet: The breast cancer cell line MCF-7 (ATCC HTB 22) and healthy mouse fibroblast cell line L929 (ATCC CCL-1) were used in this study.

Techniques: Membrane