Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: The expression of FECH in (A) T24 and (B) MGH-U3 cells treated to silence ferrochelatase. Cell toxicity of (C) T24 and (D) MGH-U3 cells with silenced FECH treated with 5-aminolevulinic acid. *P<0.05, **P<0.01 and ***P<0.001. FECH, ferrochelatase; NC, negative control; si-, siRNA; 5-ALA, 5-aminolevulinic acid.

Article Snippet: Following blocking in Tris-buffered saline (pH 7.6) that contained 5% skim milk for 1 h at room temperature, the membrane was incubated overnight at 4°C with an anti-ferritin heavy chain rabbit monoclonal antibody (product code ab75972; dilution 1:1,000; Abcam), an anti-ferrochelatase mouse monoclonal antibody (cat. no. sc-377377; dilution 1:100; Santa Cruz Biotechnology, Inc.) or an anti-actin mouse monoclonal antibody (cat. no. A2066; dilution 1:3,000; Sigma-Aldrich; Merck KGaA), which was used as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated goat anti-mouse IgG (cat. no. SA00001-1; dilution 1:10,000) or anti-rabbit IgG antibody (cat. no. SA00001-2; dilution 1:10,000; both from ProteinTech Group, Inc.) at room temperature.

Techniques: Expressing, Negative Control

Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: (A) Images and (B) intensity of mitochondrial Fe 2+ in T24 cells with silenced ferrochelatase treated with 1 mM 5-aminolevulinic acid. *P<0.05 and ***P<0.001. si-, siRNA; NC, negative control; FECH, ferrochelatase; 5-ALA, 5-aminolevulinic acid; n.s, not significant.

Article Snippet: Following blocking in Tris-buffered saline (pH 7.6) that contained 5% skim milk for 1 h at room temperature, the membrane was incubated overnight at 4°C with an anti-ferritin heavy chain rabbit monoclonal antibody (product code ab75972; dilution 1:1,000; Abcam), an anti-ferrochelatase mouse monoclonal antibody (cat. no. sc-377377; dilution 1:100; Santa Cruz Biotechnology, Inc.) or an anti-actin mouse monoclonal antibody (cat. no. A2066; dilution 1:3,000; Sigma-Aldrich; Merck KGaA), which was used as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated goat anti-mouse IgG (cat. no. SA00001-1; dilution 1:10,000) or anti-rabbit IgG antibody (cat. no. SA00001-2; dilution 1:10,000; both from ProteinTech Group, Inc.) at room temperature.

Techniques: Negative Control

Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: (A) Images and (B) intensity of mitochondrial Fe 2+ in MGH-U3 cells with silenced ferrochelatase treated with 1 mM 5-aminolevulinic acid. *P<0.05 and ***P<0.001. si-, siRNA; NC, negative control; FECH, ferrochelatase; 5-ALA, 5-aminolevulinic acid; n.s, not significant.

Article Snippet: Following blocking in Tris-buffered saline (pH 7.6) that contained 5% skim milk for 1 h at room temperature, the membrane was incubated overnight at 4°C with an anti-ferritin heavy chain rabbit monoclonal antibody (product code ab75972; dilution 1:1,000; Abcam), an anti-ferrochelatase mouse monoclonal antibody (cat. no. sc-377377; dilution 1:100; Santa Cruz Biotechnology, Inc.) or an anti-actin mouse monoclonal antibody (cat. no. A2066; dilution 1:3,000; Sigma-Aldrich; Merck KGaA), which was used as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated goat anti-mouse IgG (cat. no. SA00001-1; dilution 1:10,000) or anti-rabbit IgG antibody (cat. no. SA00001-2; dilution 1:10,000; both from ProteinTech Group, Inc.) at room temperature.

Techniques: Negative Control

Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: Images and expression of ferritin in (A and B) T24 and (C and D) MGH-U3 cells with silenced ferrochelatase treated with 5-aminolevulinic acid. *P<0.05, **P<0.01 and ***P<0.001. si-, siRNA; NC, negative control; FECH, ferrochelatase; 5-ALA, 5-aminolevulinic acid; n.s, not significant.

Article Snippet: Following blocking in Tris-buffered saline (pH 7.6) that contained 5% skim milk for 1 h at room temperature, the membrane was incubated overnight at 4°C with an anti-ferritin heavy chain rabbit monoclonal antibody (product code ab75972; dilution 1:1,000; Abcam), an anti-ferrochelatase mouse monoclonal antibody (cat. no. sc-377377; dilution 1:100; Santa Cruz Biotechnology, Inc.) or an anti-actin mouse monoclonal antibody (cat. no. A2066; dilution 1:3,000; Sigma-Aldrich; Merck KGaA), which was used as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated goat anti-mouse IgG (cat. no. SA00001-1; dilution 1:10,000) or anti-rabbit IgG antibody (cat. no. SA00001-2; dilution 1:10,000; both from ProteinTech Group, Inc.) at room temperature.

Techniques: Expressing, Negative Control

Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: (A) Images of the cell cycle and (B) percentage of the S phase in T24 cells with silenced ferrochelatase treated with 5-aminolevulinic acid. *P<0.05 and **P<0.01. si-, siRNA; NC, negative control; FECH, ferrochelatase; 5-ALA, 5-aminolevulinic acid; n.s, not significant.

Article Snippet: Following blocking in Tris-buffered saline (pH 7.6) that contained 5% skim milk for 1 h at room temperature, the membrane was incubated overnight at 4°C with an anti-ferritin heavy chain rabbit monoclonal antibody (product code ab75972; dilution 1:1,000; Abcam), an anti-ferrochelatase mouse monoclonal antibody (cat. no. sc-377377; dilution 1:100; Santa Cruz Biotechnology, Inc.) or an anti-actin mouse monoclonal antibody (cat. no. A2066; dilution 1:3,000; Sigma-Aldrich; Merck KGaA), which was used as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated goat anti-mouse IgG (cat. no. SA00001-1; dilution 1:10,000) or anti-rabbit IgG antibody (cat. no. SA00001-2; dilution 1:10,000; both from ProteinTech Group, Inc.) at room temperature.

Techniques: Negative Control

Journal: Oncology Reports

Article Title: 5-Aminolevurinic acid inhibits the proliferation of bladder cancer cells by activating heme synthesis

doi: 10.3892/or.2022.8401

Figure Lengend Snippet: (A) Images of the cell cycle and (B) percentage of the S phase in MGH-U3 cells with silenced ferrochelatase treated with 5-aminolevulinic acid. *P<0.05 and **P<0.01. si-, siRNA; NC, negative control; FECH, ferrochelatase; 5-ALA, 5-aminolevulinic acid; n.s, not significant.

Article Snippet: Following blocking in Tris-buffered saline (pH 7.6) that contained 5% skim milk for 1 h at room temperature, the membrane was incubated overnight at 4°C with an anti-ferritin heavy chain rabbit monoclonal antibody (product code ab75972; dilution 1:1,000; Abcam), an anti-ferrochelatase mouse monoclonal antibody (cat. no. sc-377377; dilution 1:100; Santa Cruz Biotechnology, Inc.) or an anti-actin mouse monoclonal antibody (cat. no. A2066; dilution 1:3,000; Sigma-Aldrich; Merck KGaA), which was used as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated goat anti-mouse IgG (cat. no. SA00001-1; dilution 1:10,000) or anti-rabbit IgG antibody (cat. no. SA00001-2; dilution 1:10,000; both from ProteinTech Group, Inc.) at room temperature.

Techniques: Negative Control