Journal: International Journal of Molecular Sciences

Article Title: Expression of Alternative Splice Variants of 6-Phosphofructo-2-kinase/Fructose-2,6-bisphosphatase-4 in Normoxic and Hypoxic Melanoma Cells

doi: 10.3390/ijms22168848

Figure Lengend Snippet: The response of malignant melanoma cells to low oxygen concentration. ( A ) Upper panel: Malignant melanoma lines, WM115 and WM266-4, were cultured with 100 µM pimonidazole (hypoxyprobe) in normoxic (N) and hypoxic conditions (H) for 16 h. The formation of pimonidazole–protein adducts were detected using Western Blot. Lower panel: Ponceau S stained membrane is shown as an internal control for equal protein loading. ( B ) Upper panels: Melanoma cells were cultured for 16 h in normoxia and hypoxia. Then HIF-1 alpha subunit accumulation was verified using the Western Blot. β-actin is shown as an internal control for equal loading. Lower panels: Densitometry analysis of Western Blot bands intensity normalized to β-actin. Relative densitometry value is the average of four independent experiments. The mean ± SEM is shown. Student’s t -test was used to evaluate the influence of hypoxia on HIF-1 alpha subunit stabilization. * p < 0.05 by Student’s t -test, ** p < 0.01 by Student’s t -test, p -value between 0.05 and 0.1 by Student’s t -test was given as an indication of the trend. ( C ) CAIX and PFKFB4 expression was analyzed by RT-qPCR in both melanoma cell lines under hypoxic and normoxic conditions. Expression data for each transcript was normalized to that for the reference gene TBP. Means ± SEM of at least five independent experiments are presented relative to expression in normoxic controls. The Student t -test was used to evaluate the differences between normoxic and hypoxic expression of CAIX and PFKFB4 . ** p < 0.01 by Student’s t -test. ( D ) Upper panels: Melanoma cells were cultured for 16 h in normoxia and hypoxia. Then CAIX and PFKFB4 expression was verified using the Western Blot. β-actin is shown as an internal control for equal loading. Lower panels: Densitometry analysis of Western Blot bands intensity normalized to β-actin. Each relative densitometry value is the average of at least four independent experiments. The mean ± SEM is shown. Studen’st t -test was used to evaluate the influence of hypoxia on CAIX and PFKFB4 expression. * p < 0.05 by Student’s t -test, ** p < 0.01 by Student’s t -test, p -value between 0.05 and 0.1 by Student’s t -test was given as an indication of the trend.

Article Snippet: Two human melanoma cell lines WM115 and WM266-4 were obtained from Rockland Immunochemicals (Limerick, PA, USA).

Techniques: Concentration Assay, Cell Culture, Western Blot, Staining, Membrane, Control, Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Biocompatibility of different concentrations (μg/mL) of the two TiO 2 NPs samples: TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or pH 5.5 (HT2), as shown by cell viability, lactate dehydrogenase (LDH), and nitric oxide (NO) release assays after 24 and 72 h exposure on normal skin ( a , c , e ) and lung ( b , d , f ) fibroblasts. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05 and ** p < 0.01 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Relative levels of catalase specific activity in normal skin ( a ) and lung ( b ) fibroblasts exposed to different concentrations (31.25, 62.5 and 125 μg/mL) of TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or 5.5 (HT2) for 24 and 72 h. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05 and ** p < 0.01 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Activity Assay, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Glutathione-dependent enzymes’ activities and glutathione level in normal skin ( a , c , e ) and lung ( b , d , f ) fibroblasts exposed to different concentrations (31.25, 62.5 and 125 μg/mL) of TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or 5.5 (HT2) for 24 and 72 h. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Malondialdehyde (MDA) levels in normal skin ( a ); and lung ( b ) fibroblasts exposed to different concentrations (31.25, 62.5, and 125 μg/mL) of TiO 2 -1% Fe–N NPs co-precipitated at pH 8.5 (HT1) or 5.5 (HT2) for 24 and 72 h. Results are expressed as the mean ± standard deviation (SD) ( n = 3) and represented relative to the untreated cells (control). * p < 0.05 and ** p < 0.01 compared to control.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Interaction of New-Developed TiO 2 -Based Photocatalytic Nanoparticles with Pathogenic Microorganisms and Human Dermal and Pulmonary Fibroblasts

doi: 10.3390/ijms18020249

Figure Lengend Snippet: Actin cytoskeleton organization of skin ( a ); and lung ( b ) fibroblasts after 24 and 72 h of incubation with different concentrations (μg/mL) of the two TiO 2 NPs samples: TiO 2 -1% Fe–N co-precipitated at pH 8.5 (HT1) or pH 5.5 (HT2). F-actin (green) was labeled with phalloidin-fluorescein isothiocyanate (FITC). Scale bar: 100 μm.

Article Snippet: Normal human fibroblasts from skin (CCD-1070Sk cell line, ATCC Cat. No. CRL-2091) and from lung (MRC-5 cell line, ATCC Cat. No. CCL-171) were grown in complete Eagle’s minimum essential medium (MEM; Gibco/Invitrogen, Carlsbad, CA, USA) with the addition of 10% fetal bovine serum (FBS; Gibco/Invitrogen, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere with 5% CO 2 .

Techniques: Incubation, Labeling