human nrf2 small interfering (si) rna (sinrf2) sc-3703  (Santa Cruz Biotechnology)

Journal: Clinical and Translational Radiation Oncology

Article Title: Differential regulation of radioadaptation by quercetin between human normal and cancer cells

doi: 10.1016/j.ctro.2025.101099

Figure Lengend Snippet: Quercetin restores NRF2 nuclear translocation in radioadapted MCF10A cells. Representative immunofluorescence images of NRF2 (green) and DAPI (blue) and quantification of the nuclear-to-cytoplasmic NRF2 fluorescence ratio in MCF10A cells 24 h after 5 Gy irradiation with or without prior LDRT and quercetin treatment (Scale bar: 10 μm). Data is shown as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were incubated with rabbit anti-human NRF2 primary antibody (1:200; Proteintech, 16396–1-AP), and detection was performed using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Irradiation

Journal: Science Advances

Article Title: Targeting HMGB2 acts as dual immunomodulator by bolstering CD8 + T cell function and inhibiting tumor growth in hepatocellular carcinoma

doi: 10.1126/sciadv.ads8597

Figure Lengend Snippet: ( A ) Location of differential accessible ATAC-seq peaks in NC and Hmgb2 -cKO CD8 + T cells. 3′UTR, 3′ untranslated region; 5′UTR, 5′ untranslated region. ( B ) Chromatin accessibility changes of genes associated with effector function and mitochondrial transcription factors. ( C ) The correlation between Hmgb2 , Tfam , and Tfb1m expression in CD8 + T cells from murine scRNA-seq data. ( D ) mRNA levels of Keap1 and Nfe2l2 in NC and Hmgb2 -cKO CD8 + T cells ( n = 6). ( E ) mRNA levels of ARE genes in NC and Hmgb2 -cKO CD8 + T cells ( n = 3). ( F ) Protein changes of KEAP1 and NRF2 after Hmgb2 knockout. ( G ) The ubiquitination of NRF2 after Hmgb2 knockout. IB, immunoblot. ( H ) Protein changes of KEAP1 and NRF2 after stimulation of mouse recombinant HMGB2 protein and IN-1. NC CD8 + T cells were treated with recombinant HMGB2 protein (500 ng/ml) and/or IN-1 (10 μM) for 48 hours. ( I ) The ubiquitination of NRF2 after cell treatment as in (H). ( J ) Immunofluorescence staining of spontaneous HCC tissues in NC and Hmgb2 -cKO mice. NRF2 + KEAP1 − CD8 + T cells were labeled. Scale bars, 20 μm (left) and 10 μm (right). HP, high power field. ( K ) Seahorse extracellular flux analysis of OCR in different CD8 + T cell groups ( n = 6 to 8) after cell treatment as in (H). ( L ) Quantification of seahorse extracellular flux analysis of OCR of different CD8 + T cells as in (K). Data are presented as the means ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Pearson test for (C) Student’s t test for (D), (E), and (J). Two-way ANOVA test for (L).

Article Snippet: Anti-human/mouse NRF2 , 16396-1-AP , Proteintech, China.

Techniques: Expressing, Knock-Out, Ubiquitin Proteomics, Western Blot, Recombinant, Immunofluorescence, Staining, Labeling

Journal: Science Advances

Article Title: Targeting HMGB2 acts as dual immunomodulator by bolstering CD8 + T cell function and inhibiting tumor growth in hepatocellular carcinoma

doi: 10.1126/sciadv.ads8597

Figure Lengend Snippet: Markers and article numbers of antibodies. PE, phycoerythrin; FITC, fluorescein isothiocyanate; APC, antigen-presenting cell; HRP, horseradish peroxidase; mAb, monoclonal antibody.

Article Snippet: Anti-human/mouse NRF2 , 16396-1-AP , Proteintech, China.

Techniques: Ubiquitin Proteomics, Purification, In Vivo

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) TCGA data analysis showing the correlation between SDCBP mRNA and BACH1 mRNA expression in GSE142102 ( n = 226) dataset of TNBC patients (Pearson correlation coefficient r = 0.3245, P < 0.0001). ( B ) TCGA data analysis showing the correlation between SDCBP mRNA and BACH1 mRNA expression in GSE103091 ( n = 238) dataset of TNBC patients (Pearson correlation coefficient r = 0.2120, P < 0.001). ( C ) Western blot showing SDCBP, BACH1, and HO-1 protein expression in MDA-MB-231, MDA-MB-468, Hs578T, MCF-7, and T47D cells. ( D ) The expression levels of SDCBP and BACH1 protein in Fig. EV1C were quantified using densitometry and normalized to the housekeeping protein α-tubulin ( n = 3). ( E ) Real-time qPCR showing SDCBP and BACH1 mRNA expression in MDA-MB-231, MDA-MB-468, Hs578T, MCF-7, and T47D cells ( n = 3). Quantitative data were normalized to β-actin expression. ( F ) Western blot showing SDCBP and HO-1 protein expression in MDA-MB-231 cells transfected with scramble or BACH1 siRNA. ( G ) Left, western blot showing the protein expression of SDCBP in the scramble and in several SDCBP-KO MDA-MB-231 subclones generated using CRISPR-Cas9 system; Right, real-time qPCR showing the SDCBP mRNA expression in scramble and in SDCBP-KO MDA-MB-231 subclones ( n = 3). ( H ) Real-time qPCR showing the mRNA expression of BACH1 in MDA-MB-231 cells, in scramble and in SDCBP-KO MDA-MB-231 subclone#2 and subclone#12 ( n = 3). ( I ) Immunofluorescence staining was used to visualize SDCBP (green color) and BACH1 (red color) in scramble and in SDCBP-KO MDA-MB-231 cells. DAPI (blue color) was used to stain the nucleus ( n = 3); Representative confocal immunofluorescence images are shown. Scale bar = 20 µm. ( J ) Western blot showing BACH1 and HO-1 protein expression in 4T1 cells infected with scramble or adenoviral SDCBP shRNA. ( K ) Real-time qPCR showing the mRNA expression of BACH1-regulated antioxidant genes ( HMOX1, NQO1 , and GLCL ) in 4T1 cells transfected with scramble or SDCBP siRNA ( n = 3); mRNA expression of KEAP1 was the negative control. Data are expressed as the mean ± SEM and analyzed using one-way ANOVA ( D , E , G , H ) or two-way ANOVA ( K ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated.

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Expressing, Western Blot, Transfection, Generated, CRISPR, Immunofluorescence, Staining, Infection, shRNA, Negative Control

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Immunohistochemistry staining against the SDCBP and BACH1 protein in human TNBC-derived tissue microarray sections ( n = 78). Representative images showing the co-expression of SDCBP and BACH1 in the same section. Normal breast cancer tissues were considered as the negative control. Scale bar = 20 µm. ( B ) Pearson correlation coefficient (r = 0.5772, P < 0.0001) between SDCBP and BACH1 expression in ( A ). ( C ) Western blot showing BACH1 and HO-1 protein expression in Hs578T cells transfected with control vector or Myc-SDCBP-expressing vector. ( D ) Real-time qPCR showing BACH1 mRNA expression in Hs578T cells transfected with a control vector or a Myc-SDCBP-expressing vector ( n = 3). ( E ) Real-time qPCR showing the mRNA expression of BACH1-regulated antioxidant genes ( HMOX1 and NQO1 ) and BACH1-regulated metastatic genes ( HK2, MMP1 , and CXCR4 ) in Hs578T cells transfected with control vector or Myc-SDCBP-expressing vector ( n = 3). ( F ) Western blot showing BACH1 protein expression in MDA-MB-231 infected with lentiviral scramble or SDCBP shRNA. ( G ) Real-time qPCR showing BACH1 mRNA expression in MDA-MB-231 infected with lentiviral scramble or SDCBP shRNA ( n = 3). ( H ) Left, Representative images of immunofluorescence staining to visualize SDCBP ( green color ) and BACH1 ( red color ) expression in MDA-MB-231 cells transfected with a scramble siRNA or SDCBP siRNA. DAPI ( blue color) was used to stain the nucleus ( n = 3); Scale bar = 50 µm. Right, fluorescence levels of SDCBP and BACH1 were quantified based on their spectral densities. ( I ) Real-time qPCR showing the mRNA expression of BACH1-regulated antioxidant genes ( HMOX1 and NQO1 ) and BACH1-regulated metastatic genes ( HK2, MMP1, MMP13 , and CXCR4 ) in MDA-MB-231 cells transfected with scramble siRNA or SDCBP siRNA ( n = 3). ( J ) Western blot showing SDCBP and BACH1 protein expression in scramble control and two SDCBP-KO MDA-MB-231 clones (KO#2, KO#12) generated using CRISPR-Cas9 system ( n = 3). ( K ) Real-time qPCR showing the mRNA expression of BACH1-regulated metastatic genes ( HK2 , MMP1 , CXCR4, GAPDH , and VEGF ) in scramble control and SDCBP-KO MDA-MB-231 cells. ( L ) The reconstitution of SDCBP recovers BACH1 protein expression in SDCBP-KO MDA-MB-231 cells. Western blot showing BACH1 protein expression in scramble control and SDCBP-KO MDA-MB-231 cells transfected with control vector or Myc-SDCBP-expressing vector. The arrows indicate the endogenous (Endo) and exogenous (Exo) SDCBP. Data are expressed as the mean ± SEM and analyzed using two-tailed Student’s t test with Welch’s correction ( D , E , G , I ), two-way ANOVA ( H ), or one-way ANOVA ( K ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated. .

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Immunohistochemistry, Staining, Derivative Assay, Microarray, Expressing, Negative Control, Western Blot, Transfection, Control, Plasmid Preparation, Infection, shRNA, Immunofluorescence, Fluorescence, Clone Assay, Generated, CRISPR, Two Tailed Test

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Western blot showing BACH1 protein expression in Hs578T cells transfected with Myc-SDCBP-expressing vector with or without BACH1 siRNA. ( B ) Colony formation of Hs578T cells transfected with Myc-SDCBP-expressing vector with or without BACH1 siRNA ( n = 3); See also Fig. . Migration of Hs578T cells transfected with Myc-SDCBP-expressing vector with or without BACH1 siRNA ( n = 3); See also Fig. . ( D ) Western blot showing SDCBP and Flag-BACH1 protein expression in MDA-MB-231 cells transfected with SDCBP siRNA with or without a Flag-BACH1-expressing vector. Cell proliferation of MDA-MB-231 cells transfected with SDCBP siRNA with or without Flag-BACH1-expressing vector ( n = 3). ( F ) Colony formation of MDA-MB-231 cells transfected with SDCBP siRNA with or without Flag-BACH1-expressing vector ( n = 3); See also Fig. . ( G ) Wound closure of scratched MDA-MB-231 cells transfected with SDCBP siRNA with or without a Flag-BACH1-expressing vector ( n = 3); See also Fig. . ( H ) Schematic of various SDCBP mutant constructs generated using the Myc-SDCBP plasmid. ( I ) Left, western blot showing BACH1 protein expression in Hs578T cells transfected with the indicted SDCBP constructs. Right, quantification of BACH1 levels using densitometry ( n = 3). ( J ) Top, schematic of the PDZ1 construct. Bottom, western blot showing BACH1 protein expression in Hs578T cells transfected with Myc-SDCBP or Myc-SDCBP_PDZ1 plasmid. ( K ) Migration of Hs578T cells transfected with Myc-SDCBP, Myc-SDCBP_Δ4, or Myc-SDCBP_PDZ1 plasmid ( n = 3); See also Fig. . ( L ) Colony formation of Hs578T cells transfected with Myc-SDCBP, Myc-SDCBP_Δ4, or Myc-SDCBP_PDZ1 plasmid ( n = 3); See also Fig. . ( M ) Tumor volumes from athymic BALB/c nude mice 6 weeks after mammary fat-pad injection of the scramble control or SDCBP-KO MDA-MB-231 cells (1 × 10 5 cells/mouse; n = 5 or 7 mice/group). ( N ) Tumor weights in Fig. 2M ( n = 7 mice/group); See also Fig. . ( O ) Tumor volumes from athymic BALB/c nude mice 25 days after mammary fat-pad injection of the scramble control, SDCBP-KO MDA-MB-231 cells, and SDCBP-KO MDA-MB-231 cells stably transfected with Flag-SDCBP (1 × 10 5 cells/mouse; n = 5 mice/group); See also Fig. – , . Data are expressed as the mean ± SEM and analyzed using one-way ANOVA ( B , , I , K , L , N ), two-tailed Student’s t test ( , F , O ), or two-way ANOVA ( G , M ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated. .

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Migration, Mutagenesis, Construct, Generated, Injection, Control, Stable Transfection, Two Tailed Test

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Representative images of colony formation in Fig. . ( B ) Representative images of the migrated cells in Fig. . Scale bar = 200 µm. ( C ) Representative images of colony formation in Fig. . ( D ) Representative images of wounding migration in Fig. . Scale bar = 200 µm. ( E ) Cell proliferation of MDA-MB-231 cells transfected with scramble or BACH1 siRNA. Cell proliferation was estimated by an automatic cell counter at the indicated time points ( n = 3). ( F ) Colony formation of MDA-MB-231 cells transfected with scramble or BACH1 siRNA. The clonogenic ability was assessed and quantified based on the absorbance at 600 nm and normalized to the control ( n = 3). ( G ) Migration of MDA-MB-231 cells transfected with scramble or BACH1 siRNA. The number of migrated cells were counted and expressed as percentages ( n = 3). ( H ) Representative images of the migrated cells in Fig. . Scale bar = 500 µm. ( I ) Representative images of colony formation in Fig. . Scale bar = 1000 µm. Data are expressed as the mean ± SEM and analyzed using two-tailed Student’s t test with Welch’s correction ( E – G ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated.

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Migration, Transfection, Control, Two Tailed Test

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with SDCBP siRNA with or without MG132 proteasome inhibitor. ( B ) Left, western blot showing BACH1 protein expression in Hs578T cells transfected with control vector or Myc-SDCBP-expressing vector in the presence of CHX protein synthesis inhibitor at various time points. Right, quantification of BACH1 protein levels using densitometry ( n = 3). ( C ) Left, western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with scramble or FBXO22 siRNA in the presence of CHX protein synthesis inhibitor at various time points. Right, quantification of BACH1 protein levels using densitometry ( n = 3). ( D ) Western blot showing BACH1, FBXO22, and SDCBP protein expression in MDA-MB-231 cells transfected with scramble or FBXO22 siRNA. ( E ) Western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with HA-FBXO22-expressiong vector with or without Myc-SDCBP-expressing vector. ( F ) In vitro ubiquitylation assay of the recombinant human BACH1 protein mediated by the FBXO22 complex. Active recombinant human UbcH5a protein was used as the E2 ubiquitin-conjugating enzyme for FBXO22 complex-mediated BACH1 degradative polyubiquitylation. ( G ) In vivo ubiquitylation assay showing the decrease in the K48-linked polyubiquitylation of BACH1 by SDCBP overexpression in HEK293 cells transfected with the indicated plasmids. ( H ) In vivo ubiquitylation assay showing the increase in the K48-linked polyubiquitylation of BACH1 by SDCBP KD in MDA-MB-231 cells transfected with the indicated plasmids. ( I ) In vitro ubiquitylation assay showing the inhibitory effect of SDCBP on the polyubiquitylation of BACH1 mediated by the FBXO22 complex. Active recombinant human protein UbcH5a and immunocomplex FBXO22 were added as described above. Recombinant human BACH1 and recombinant human SDCBP proteins were added at ratios of 1:1 (+) and 1:2 (++). Data are expressed as the mean ± SEM and analyzed using two-tailed Student’s t test with Welch’s correction ( B , C ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated. .

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Western Blot, Expressing, Transfection, Control, Plasmid Preparation, In Vitro, Ubiquitin Assay, Recombinant, Ubiquitin Proteomics, In Vivo, Over Expression, Two Tailed Test

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Left, Western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with control vector or Myc-SDCBP-expressing vector in the presence of CHX protein synthesis inhibitor at various time points. Right, quantification of BACH1 protein levels using densitometry ( n = 3). ( B ) Free heme level in Hs578T cells transfected with control vector or Myc-SDCBP-expressing vector ( n = 3). ( C ) Free heme level in scramble and in SDCBP - KO MDA-MB-231 cells ( n = 3). ( D ) Western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with control vector or NRF2 (encoded by NFE2L2 )-expressing vector. HO-1 protein expression was considered as the positive control. ( E ) Western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with a control or a HO-1 (encoded by HMOX1 )-expressing vector. ( F ) Western blot showing BACH1 protein expression in Hs578T cells transfected with scramble or HO-1 siRNA. ( G ) Western blot showing BACH1 protein expression in MDA-MB-231 cells transfected with the HO-1 or the catalytic inactive HO-1 mutant (H25A) plasmid. ( H ) Western blot showing BACH1 and SDCBP protein expression in MDA-MB-231 cells transfected with scramble or HOIL1 siRNA. ( I ) Western blot showing endogenous FBXO22 protein expression in several breast cancer cells. ( J ) Immunoprecipitation showing the interaction of BACH1 with FBXO22 in HEK293 cells transfected with the indicated plasmids. An arrow indicates the specific signal for HA-FBXO22. ns: none specific. ( K ) In vivo ubiquitylation assay showing the increase in the polyubiquitylation of BACH1 by FBXO22 overexpression in HEK293 cells transfected with the indicated plasmids. ( L ) In vivo ubiquitylation assay showing the increase in the polyubiquitylation of BACH1 by FBXO22 overexpression in MDA-MB-231 cells transfected with the indicated plasmids. ( M ) In vivo ubiquitylation assay showing the decrease in FBXO22-mediated polyubiquitylation of BACH1 by SDCBP overexpression in HEK293 cells transfected with the indicated plasmids. Data are expressed as the mean ± SEM and analyzed using two-tailed Student’s t test with Welch’s correction ( A – C ). All experiments were repeated at least three times unless otherwise indicated. P values less than 0.05 were considered statistically significant.

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Western Blot, Expressing, Transfection, Control, Plasmid Preparation, Positive Control, Mutagenesis, Immunoprecipitation, In Vivo, Ubiquitin Assay, Over Expression, Two Tailed Test

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Immunoprecipitation showing the interaction of FBXO22 with SDCBP in HEK293 cells transfected with the indicated plasmids. An arrow indicates the specific signal for HA-FBXO22. ( B ) Immunoprecipitation showing the interaction of SDCBP with FBXO22 in HEK293 cells transfected with the indicated plasmids. An arrow indicates the specific signal for HA-FBXO22. ( C ) Co-immunoprecipitation showing the interaction of FBXO22 with BACH1 in HEK293 cells with or without SDCBP after the indicated transfections. ( D ) Schematic of experimental design to investigate the assembly of SCF FBXO22 –BACH1 complex via His Pull-down assay and endogenous IP assay in Fig. D– . ( E ) His-pulldown assay showing the interaction of FBXO22 with SKP1 in HEK293 cells with control vector or Myc-SDCBP-expressing vector after the indicated transfections. See also Appendix Fig. S . ( F ) Western blot showing BACH1, PTEN, and PD-L1 protein expression in scramble and in SDCBP-KO MDA-MB-231 cells. Western blot showing BACH1, PTEN, and PD-L1 protein expression in A549 cells transfected with scramble or SDCBP siRNA. ( H ) Western blot showing BACH1 and PD-L1 protein expression in NCI-H1299 cells transfected with scramble or SDCBP siRNA. ( I ) Western blot showing BACH1, PTEN, and PD-L1 protein expression in Hs578T cells transfected with control vector or Myc-SDCBP-expressing vector. ( J ) In vivo ubiquitylation assay showing the inhibitory effect of SDCBP on SCF FBXO22 -mediated K48-linked polyubiquitylation of BACH1 in HEK293 cells transfected with the indicated plasmids.

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Immunoprecipitation, Transfection, Pull Down Assay, Control, Plasmid Preparation, Expressing, Western Blot, In Vivo, Ubiquitin Assay

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Crosslink immunoprecipitation showing an interaction of FBXO22 with SDCBP in Hs578T cells transfected with Myc-SDCBP-expressing vector or Myc-SDCBP_Δ4-expressing vector. Schematic showing the FBXO22 mutant constructs generated using the HA-FBXO22 plasmid. ( C ) Immunoprecipitation showing SDCBP interactions with FBXO22 and its mutant constructs in HEK293 cells transfected with the indicated plasmids. ns indicates non-specific bands. See also Fig. , . ( D ) His-Pulldown assay showing the effect of SDCBP on SKP1-CUL1-FBXO22 complex formation after the indicated transfections in HEK293 cells. See also Appendix Fig. S . ( E ) His-Pulldown assay showing the effect of SDCBP KO on the SCF FBXO22 –BACH1 complex formation in the scramble control and SDCBP-KO MDA-MB-231 cells transfected with control vector or His-SKP1-expressing vector. See also Appendix Fig. S . ( F ) Immunoprecipitation showing the effect of SDCBP KD on the SCF FBXO22 –BACH1 complex formation in MDA-MB-231 cells transfected with scramble or SDCBP siRNA. ( G ) Immunoprecipitation showing the effect of SDCBP overexpression on the SCF FBXO22 –BACH1 complex formation in Hs578T cells transfected with control vector or Myc-SDCBP-overexpressing vector. ( H ) Immunoprecipitation showing the effect of SDCBP PDZ1 domain on the SCF FBXO22 –BACH1 complex formation in Hs578T cells transfected with a control vector or a Myc-SDCBP-PDZ1-overexpressing vector. .

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Immunoprecipitation, Transfection, Expressing, Plasmid Preparation, Mutagenesis, Construct, Generated, Control, Over Expression

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Real-time qPCR showing the mRNA expression of BACH1-regulated ETC genes ( NDUFA4 , NDUFA4L2 , NDUFC2 , and COX6B2 ) in scramble and SDCBP-KO MDA-MB-231 cells ( n = 3); See also Appendix Fig. S , . Real-time qPCR showing the mRNA expression of NDUFA4 and COX6B2 in scramble, SDCBP-KO MDA-MB-231 cells, and SDCBP-KO MDA-MB-231 cells transfected with SDCBP ( n = 3). ( C ) Western blots showing the expression of mitochondrial proteins NDUFA4 and COX6B2 in MDA-MB-231 cells transfected with SDCBP siRNA in the presence or absence of FLAG-BACH1-expressing vector. ( D ) ChIP-qPCR showing BACH1 enrichments in the promoter regions of NDUFA4 and COX6B2 in the scramble and SDCBP-KO MDA-MB-231 cells. Quantitative data were normalized to IgG binding expression ( n = 3); See also Appendix Fig. S . ( E ) Left, flow cytometry histogram showing the mitochondrial membrane potentials using TMRE (tetramethylrhodamine ethyl ester) staining in MDA-MB-231 cells transfected with a scramble or SDCBP siRNA after the indicated treatments. FCCP (trifluoromethoxy carbonylcyanide phenylhydrazone). Right, quantification of TMRE fluorescence intensity ( n = 3). ( F ) Left, immunofluorescence staining and confocal imaging of the fluorescent signals for TMRE (orange-red color) in the scramble control and SDCBP-KO MDA-MB-231 cells after incubation with TMRE. DAPI (blue color) was used to stain the nucleus ( n = 7); Representative confocal images are shown; scale bars = 20 µm and 5 µm. Right, fluorescent levels of the TMRE were quantified based on their spectral densities. ( G ) Representative images of immunohistochemistry staining against the NDUFA4, BACH1, and SDCBP proteins showing a negative correlation between the expression of NDUFA4 and SDCBP in the same sections of TNBC tumor tissues. Scale bar = 20 µm. ( H ) Pearson correlation coefficients between SDCBP and NDUFA4 protein expression ( n = 64), and between BACH1 and NDUFA4 protein expression ( n = 60) in Fig. 5G. Data are expressed as the mean ± SEM and analyzed using two-way ANOVA ( A , D , F ), two-tailed Student’s t test , or one-way ANOVA ( E ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated. .

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Expressing, Transfection, Western Blot, Plasmid Preparation, ChIP-qPCR, Binding Assay, Flow Cytometry, Membrane, Staining, Fluorescence, Immunofluorescence, Imaging, Control, Incubation, Immunohistochemistry, Two Tailed Test

Journal: The EMBO Journal

Article Title: SDCBP/Syntenin-1 stabilizes BACH1 by disassembling the SCF FBXO22 –BACH1 complex in triple-negative breast cancer

doi: 10.1038/s44318-025-00440-1

Figure Lengend Snippet: ( A ) Cell proliferation of MDA-MB-231 cells transfected with a scramble or SDCBP siRNA after treatment with metformin for the indicated periods of time ( n = 3). ( B ) Cell viability of MDA-MB-231 cells transfected with a scramble or SDCBP siRNA after treatment with the indicated concentrations of metformin for 96 h ( n = 5). ( C ) Colony formation for MDA-MB-231 cells transfected with a scramble or SDCBP siRNA after treatment with the indicated concentrations of metformin. Colony numbers were counted and converted to percentages by normalizing with the control groups ( n = 3). ( D – F ) Effect of the indicated treatment on 4T1 tumor growth. Tumor growth was monitored in BALB/c mice bearing 4T1 cells after mammary fat-pad injections. When the average tumor volumes reached 100 mm 3 , the mice ( n = 7 mice/group) were administered with 100 mg/kg metformin (once a day) and/or adenoviral SDCBP shRNA (1 × 10 9 PFU/mice). Black arrows indicate the day of adenoviral SDCBP shRNA injection. Final tumor volume ( E ) and weight ( F ) are shown. ( G ) Immunohistochemistry staining against SDCBP, BACH1, Ki67, NDUFA4, and COX6B2 protein in 4T1 tumors from BALB/c mice in Fig. 6A. Representative images of the IHC staining are shown. Scale bar = 50 µm for low (left) and high (right) magnification. Data are expressed as the mean ± SEM and analyzed using two-way ANOVA ( A – C ), one-way ANOVA ( E ), or two-tailed Student’s t test ( F ). P values less than 0.05 were considered statistically significant. All experiments were repeated at least three times unless otherwise indicated. .

Article Snippet: Human NRF2 siRNA , Santa Cruz Biotechnology , Cat#sc-37030.

Techniques: Transfection, Control, shRNA, Injection, Immunohistochemistry, Staining, Two Tailed Test