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a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with <t>CRISPR/Cas9-mediated</t> CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.
Cas9 Protein (80 Ng/µl), supplied by ToolGen Incorporated, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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aureobasidin a (aba) (0, 20, 80 ng/ml)  (Millipore)
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a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with <t>CRISPR/Cas9-mediated</t> CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.
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a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with <t>CRISPR/Cas9-mediated</t> CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.
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a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with <t>CRISPR/Cas9-mediated</t> CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.
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a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with <t>CRISPR/Cas9-mediated</t> CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.
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80 ng/ml vegf  (Thermo Fisher)
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Thermo Fisher 80 ng/ml vegf
a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with <t>CRISPR/Cas9-mediated</t> CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.
80 Ng/Ml Vegf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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80 ng/ml vegf - by Bioz Stars, 2026-02
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a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with CRISPR/Cas9-mediated CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.

Journal: Experimental & Molecular Medicine

Article Title: CYB5R3 functions as a tumor suppressor by inducing ER stress-mediated apoptosis in lung cancer cells via the PERK-ATF4 and IRE1α-JNK pathways

doi: 10.1038/s12276-024-01155-9

Figure Lengend Snippet: a Immunofluorescence staining of CYB5R3 (green) and calnexin (red) was carried out in H1299 cells infected with EV or CYB5R3. DAPI (blue) was used for nuclear staining. Scale bar, 20 μm. b Protein levels of ER markers in H1299 cells infected with EV or CYB5R3 for 24 h. c H1299 cells were transfected with the indicated siRNA and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. d H1299 cells were transfected with siRNA against ATF4 and infected with CYB5R3 for 48 h. Cell viability (lower panel) or immunoblot analysis (upper panel) was performed. e H1299 cells were transfected with the indicated siRNAs and infected with CYB5R3 for 48 h. A cell viability assay (left panel) and immunoblot analysis (right panel) were performed. f Protein level of CYB5R3 (upper panel) and cell viability (lower panel) in H1299 and H1703 cells with CRISPR/Cas9-mediated CYB5R3 knockout. g Protein levels of ER markers in tunicamycin-treated CYB5R3-knockout H1299 and H1703 cells. The values are presented as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01.

Article Snippet: A mixture of the sgRNA (100 ng/µl) and the Cas9 protein (80 ng/µl) (ToolGen Inc., Seoul, Korea) was injected into the cytoplasm of pronuclear-stage embryos.

Techniques: Immunofluorescence, Staining, Infection, Transfection, Viability Assay, Western Blot, CRISPR, Knock-Out