Journal: bioRxiv

Article Title: Abnormal shear stress induces ferroptosis in endothelial cells via KLF6 downregulation

doi: 10.1101/2025.09.10.675441

Figure Lengend Snippet: KLF6 regulates the UPR and mevalonate pathway. (A) Heat map of the identified TF genes downregulated by LSS; (B) Predicted KLF6-binding motifs in promoters of PERK, HSPA5, MVD, IDI1 (JASPAR database; matrix ID: MA0472.1); (C) Protein expression of KLF6 under different shear stress conditions; (D) Validation of KLF6 overexpression by Western blot (OE: overexpression, NC: negative control); (E, F) KLF6-OE largely restores the expression of PERK, BiP, MVD, and IDI1 under LSS and HSS to levels seen under MSS. Data are expressed as the mean ± standard deviation from three or more independent experiments. Statistical significance was determined using an unpaired t-test, with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Antibodies: SLC7A11 (1:1000, ab175186, Abcam), GRP78/BIP (1:1000, 66574-1-Ig, proteintech), PERK/EIF2AK3 (1:1000, 24390-1-AP, proteintech), KLF6 (1:1000, 14716-1-AP, proteintech), MVD (1:1000, 15331-1-AP proteintech) IDI1 (1:1000, 11166-2-AP, proteintech), Goat Anti-Rabbit IgG-HRP (1:10,000, ASS1006-1, Biorigin), Goat Anti-Mouse IgG-HRP (1:10,000, ASS1007-1, Biorigin).

Techniques: Binding Assay, Expressing, Shear, Biomarker Discovery, Over Expression, Western Blot, Negative Control, Standard Deviation

Journal: bioRxiv

Article Title: Abnormal shear stress induces ferroptosis in endothelial cells via KLF6 downregulation

doi: 10.1101/2025.09.10.675441

Figure Lengend Snippet: KLF6 overexpression alleviates endothelial ferroptosis and early atherosclerotic events induced by LSS and HSS. (A, E) Protein expression of SLC7A11; (B, F) ELISA results of extracellular LDH; (C, G) ELISA results of intracellular CoQ10; (D, H) Flow cytometry results of lipid peroxidation indicated by C11 BODIPY; (I, K) Fluorescent microscopic results of THP-1 cells (green label) adhering to the VSMC-EC bilayer cell architecture (The scale bar = 250 μm); (J, L) Lipid accumulation visualized by Oil Red O staining (The scale bar = 25 μm). Data are expressed as the mean ± standard deviation from three or more independent experiments. Statistical significance was determined using an unpaired t-test, with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Antibodies: SLC7A11 (1:1000, ab175186, Abcam), GRP78/BIP (1:1000, 66574-1-Ig, proteintech), PERK/EIF2AK3 (1:1000, 24390-1-AP, proteintech), KLF6 (1:1000, 14716-1-AP, proteintech), MVD (1:1000, 15331-1-AP proteintech) IDI1 (1:1000, 11166-2-AP, proteintech), Goat Anti-Rabbit IgG-HRP (1:10,000, ASS1006-1, Biorigin), Goat Anti-Mouse IgG-HRP (1:10,000, ASS1007-1, Biorigin).

Techniques: Over Expression, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Staining, Standard Deviation

Journal: bioRxiv

Article Title: Abnormal shear stress induces ferroptosis in endothelial cells via KLF6 downregulation

doi: 10.1101/2025.09.10.675441

Figure Lengend Snippet: Overview of KLF6 mediated the ferroptosis induced by abnormal shear stress

Article Snippet: Antibodies: SLC7A11 (1:1000, ab175186, Abcam), GRP78/BIP (1:1000, 66574-1-Ig, proteintech), PERK/EIF2AK3 (1:1000, 24390-1-AP, proteintech), KLF6 (1:1000, 14716-1-AP, proteintech), MVD (1:1000, 15331-1-AP proteintech) IDI1 (1:1000, 11166-2-AP, proteintech), Goat Anti-Rabbit IgG-HRP (1:10,000, ASS1006-1, Biorigin), Goat Anti-Mouse IgG-HRP (1:10,000, ASS1007-1, Biorigin).

Techniques: Shear

Journal: Animal Bioscience

Article Title: MiRNA-24 downregulates KLF6 affecting STAT3 protein expression and phosphorylation regulating melanogenesis in cashmere goat coat

doi: 10.5713/ab.24.0824

Figure Lengend Snippet: Validation of MiRNA-24 cashmere color correlation and targeting relationship. (a) Target gene screening based on miRDB and Target Scan databases. (b) Target gene GO/KEGG enrichment analysis chart. (c) The expression level of miRNA-24 in the skin of black and white Cashmere goats (** p<0.01). (d) The bacterial liquid sequencing image of the reconstructed plasmid shows the binding sequences of psiCHECK-2-KLF6-WT and psiCHECK-2-KLF6-MUT 3'UTR, marked in yellow. (e) The point mutation design of KLF6-WT and KLF6-MUT 3'UTR binding sequences, with binding sites marked in red. (f) The result of dual luciferase assay (** p<0.01). WT, wild-type; MUT, mutant-type.

Article Snippet: KLF6 Monoclonal antibody (1:2,500; Proteintech, Wuhan, China), KLF6 Polyclonal Antibody (1:2,500; Proteintech), STAT3 antibody (1:1,000; Abmart, Berkeley Heights, NJ, USA), STAT3 (phospho Ser727) antibody (1:1,000; Abmart) and β-Actin antibody (1:2,000; ZSGB, Beijing, China) were incubated with the bands for a whole night at 4°C.

Techniques: Biomarker Discovery, Expressing, Sequencing, Plasmid Preparation, Binding Assay, Mutagenesis, Luciferase

Journal: Animal Bioscience

Article Title: MiRNA-24 downregulates KLF6 affecting STAT3 protein expression and phosphorylation regulating melanogenesis in cashmere goat coat

doi: 10.5713/ab.24.0824

Figure Lengend Snippet: MiRNA-24 regulates the expression of KLF6 . (a) KLF6 is relatively expressed in the skin of black and white Cashmere goats (** p<0.01). (b) Relative protein expression and bands of KLF6 in black and white Cashmere goat skin (N = 3, ** p<0.01). (c) The relative gene expression of miRNA-24 in B16-F10 cells transfected with miRNA-24 mimics/inhibitors/NC. (d) Relative gene and protein expression of KLF6 in B16-F10 cells transfected with miRNA-24 mimics/inhibitors/ NC (N = 3, * p<0.05, ** p<0.01). (e) The protein localization and expression of KLF6 in B16-F10 cells in each group, as shown in the left figure ×40 and the right figure ×1,000. N, number of biological repeated experiments.

Article Snippet: KLF6 Monoclonal antibody (1:2,500; Proteintech, Wuhan, China), KLF6 Polyclonal Antibody (1:2,500; Proteintech), STAT3 antibody (1:1,000; Abmart, Berkeley Heights, NJ, USA), STAT3 (phospho Ser727) antibody (1:1,000; Abmart) and β-Actin antibody (1:2,000; ZSGB, Beijing, China) were incubated with the bands for a whole night at 4°C.

Techniques: Expressing, Gene Expression, Transfection

Journal: Animal Bioscience

Article Title: MiRNA-24 downregulates KLF6 affecting STAT3 protein expression and phosphorylation regulating melanogenesis in cashmere goat coat

doi: 10.5713/ab.24.0824

Figure Lengend Snippet: The mechanism by which KLF6 affects melanogenesis. (a) After silencing KLF6 (si-KLF6), the relative expression levels of KLF6 gene and protein (N = 3, *** p<0.001). (b) The mRNA levels of genes related to the melanogenesis pathway (** p<0.01, *** p<0.001). (c) The mRNA levels of STAT3 and TYR after silencing KLF6 (** p<0.01). (d) After KLF6 silencing, STAT3 protein and its phosphorylation levels (N = 3, ** p<0.01, *** p<0.001). (e) After STAT3 silencing, the expression levels of STAT3 protein and its phosphorylation (N = 3, * p<0.05). (f) The expression levels of STAT3 and TYR mRNA after STAT3 silencing (** p<0.01). (g) The protein and phosphorylation levels after STAT3 phosphorylation inhibition (N = 3, ** p<0.01). (h) The mRNA expression level of TYR after STAT3 phosphorylation inhibition (** p<0.01). N, number of biological repeated experiments.

Article Snippet: KLF6 Monoclonal antibody (1:2,500; Proteintech, Wuhan, China), KLF6 Polyclonal Antibody (1:2,500; Proteintech), STAT3 antibody (1:1,000; Abmart, Berkeley Heights, NJ, USA), STAT3 (phospho Ser727) antibody (1:1,000; Abmart) and β-Actin antibody (1:2,000; ZSGB, Beijing, China) were incubated with the bands for a whole night at 4°C.

Techniques: Expressing, Phospho-proteomics, Inhibition

Journal: Animal Bioscience

Article Title: MiRNA-24 downregulates KLF6 affecting STAT3 protein expression and phosphorylation regulating melanogenesis in cashmere goat coat

doi: 10.5713/ab.24.0824

Figure Lengend Snippet: The animal replenishment experiment. (a) The relative gene expressions of miRNA-24 and KLF6 in the AntagomiRNA-24 and PBS (control) group (** p<0.01, *** p<0.001). (b) The relative protein levels of KLF6 in the AntagomiRNA-24 and PBS group (N = 3, ** p<0.01). (c) The standard curve of melanin on the left and the melanin contents in the skin of the AntagomiRNA-24 and PBS group (*** p<0.001). PBS, phosphate-buffered saline; OD, optical density; N, number of biological repeated experiments.

Article Snippet: KLF6 Monoclonal antibody (1:2,500; Proteintech, Wuhan, China), KLF6 Polyclonal Antibody (1:2,500; Proteintech), STAT3 antibody (1:1,000; Abmart, Berkeley Heights, NJ, USA), STAT3 (phospho Ser727) antibody (1:1,000; Abmart) and β-Actin antibody (1:2,000; ZSGB, Beijing, China) were incubated with the bands for a whole night at 4°C.

Techniques: Control, Saline