Journal: Cell Proliferation

Article Title: 4‐Octyl itaconate blocks GSDMB ‐mediated pyroptosis and restricts inflammation by inactivating granzyme A

doi: 10.1111/cpr.13711

Figure Lengend Snippet: Metabolic derivative 4‐OI suppresses GSDMB‐mediated pyroptosis independent of Nrf2. (A, B) Western blot analysis of GSDMB cleavage in GSDMB‐expressing 293 T cells and (C) LDH release activity from the media when exposed to several metabolites. (D, E) Cleavage of GSDMB and (F) LDH release activity in GSDMB‐expressing 293 T cells when exposed to different concentrations of 4‐OI. (G) Representative image of live/dead assay in GSDMB‐expressing 293 T cells after electroporated with GrzA when exposed to different concentrations of 4‐OI. Live cell (green), Calcein AM. Dead cell (red), PI. Scale bar, 100 μm. Shown on the right was the number of PI+ cells. (H) Cleavage of GSDMB and (I) LDH release activity in human primary IECs after 4‐OI treatment. (J) Western blot analysis of GSDMB cleavage and (K) LDH release activity when transfected with siRNA of Nrf2. Data were displayed as mean values ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The double‐stranded siRNA targeting human GSDMB and Nrf2 were synthesised by General Biosystems (Anhui, China) Co. Ltd.

Techniques: Western Blot, Expressing, Activity Assay, Live Dead Assay, Transfection

Journal: Oncotarget

Article Title: NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction

doi: 10.18632/oncotarget.12435

Figure Lengend Snippet: ( A , B ) The effect of NRF2 mRNA expression level on the relapse free survival (A) and post progression survival (B) in 4,142 breast cancer patients was analyzed. The Kaplan-Meier plots were generated by Kaplan-Meier Plotter ( http://www.kmplot.com ). ( C – E ) The effect of NRF2 mRNA expression level on the relapse free survival of ER-negative samples (C), ER-negative and HER2-negative samples (D) or ER-negative and HER2-positive samples (E). ( F – H ) The effect of NRF2 mRNA expression level on the relapse free survival of ER-positive samples (F), ER-positive and HER2-negative samples (G) or ER-positive and HER2-positive samples (H).

Article Snippet: NRF2 siRNA, RhoA siRNA and a scramble non-targeting siRNA (siCtrl) were purchased from Biomics Biotech (Biomics Biotechnologies Co., Ltd, Nan Tong, China).

Techniques: Expressing, Generated

Journal: Oncotarget

Article Title: NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction

doi: 10.18632/oncotarget.12435

Figure Lengend Snippet: ( A , B ) NRF2 expression was effectively decreased at both mRNA (A) and protein levels (B) in the MDA-MB-231 cell line. ( C , D ) NRF2 expression was effectively decreased at both mRNA (C) and protein levels (D) in the MCF7 cell line. n = 3, bar: SD, *** P < 0.005.

Article Snippet: NRF2 siRNA, RhoA siRNA and a scramble non-targeting siRNA (siCtrl) were purchased from Biomics Biotech (Biomics Biotechnologies Co., Ltd, Nan Tong, China).

Techniques: Expressing

Journal: Oncotarget

Article Title: NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction

doi: 10.18632/oncotarget.12435

Figure Lengend Snippet: ( A , B , E , F ) qRT-PCR analysis of the mRNA expression levels of NRF2 and RhoA in MDA-MB-231 and MCF7 cells after being transfected with siRNA specifically targeting to the human NRF2 gene or RhoA gene. ( C , D , G , H ) Immunoblotting of NRF2 and RhoA in MDA-MB-231 and MCF7 cells transfected with siNrf2 or siRhoA. n = 3. ( I , J ) Relative protein expression levels were quantified using ImageJ and normalized to β-actin and then to their corresponding siCtrl -transfected cells. All data are means of three experiments, bar: SD, * P < 0.05; ** P < 0.01; *** P < 0.005.

Article Snippet: NRF2 siRNA, RhoA siRNA and a scramble non-targeting siRNA (siCtrl) were purchased from Biomics Biotech (Biomics Biotechnologies Co., Ltd, Nan Tong, China).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Western Blot

Journal: Oncotarget

Article Title: NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction

doi: 10.18632/oncotarget.12435

Figure Lengend Snippet: ( A , B ) MDA-MB-231 and MCF7 cells were transiently transfected with siNrf2 alone, or simultaneously with RhoA expressing vectors or treated with Compound 1. The protein levels of NRF2 and RhoA were determined by immunoblotting. ( C ) The intensity of each band was quantified using ImageJ and normalized to β-actin and then to their corresponding siCtrl-transfected cells. n = 3, bar: SD, *** P < 0.005. ( D – G ) Cell proliferation was measured by Ki67 immunostaining. (D, E) Cells were stained with anti-Ki67 antibodies to detect cell proliferation ability (green), and with DAPI, to detect nuclei (blue). n = 5. (F, G) Ki67 staining rate was quantified by Image J. ( H , I ) Cell growth was determined using thiazolyl blue assay at various time points. Cells were treated with siCtrl, siNrf2 alone, siNrf2 combined with RhoA expressing vectors or siRhoA combined with Compound 1. n = 10, bar: SD, * P < 0.05; ** P < 0.01; *** P < 0.005.

Article Snippet: NRF2 siRNA, RhoA siRNA and a scramble non-targeting siRNA (siCtrl) were purchased from Biomics Biotech (Biomics Biotechnologies Co., Ltd, Nan Tong, China).

Techniques: Transfection, Expressing, Western Blot, Immunostaining, Staining

Journal: Oncotarget

Article Title: NRF2 promotes breast cancer cell proliferation and metastasis by increasing RhoA/ROCK pathway signal transduction

doi: 10.18632/oncotarget.12435

Figure Lengend Snippet: ( A ) The relationship between NRF2 and ERR1 mRNA expression was retrieved from TCGA dataset using www.cbioportal.org and the correlation was analyzed by Pearson correlation and Spearman correlation. ( B , C ) The expression of ERR1 mRNA and protein levels was measured by qRT-PCR (B) and immunoblotting (C) in MCF7 cells. ( D , E ) The expression of ERR1 mRNA and protein levels was measured by qRT-PCR (D) and immunoblotting (E), respectively, in MDA-MB-231 cells. All data are means of three experiments, bar: SD, * P < 0.05; ** P < 0.01; *** P < 0.005. ( F , G ) Binding of NRF2 to the ERR1 or HO-1 promoter in MCF7 (F) and MDA-MB-231 cells (G). Anti-NRF2- or IgG-immunoprecipitated chromatin was amplified using the indicated primer pairs. Results obtained by real-time PCR are expressed relative to amplified input. n = 3. Conventional PCR products from duplicate ChIP were analyzed on agarose gels. Input was diluted 1/100 before PCR. HO-1 gene served as positive control.

Article Snippet: NRF2 siRNA, RhoA siRNA and a scramble non-targeting siRNA (siCtrl) were purchased from Biomics Biotech (Biomics Biotechnologies Co., Ltd, Nan Tong, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Immunoprecipitation, Amplification, Real-time Polymerase Chain Reaction, Positive Control