Journal: Genes & Diseases

Article Title: Autophagy-induced NR2F1 activation promotes the apoptosis of lens epithelial cells and facilitates cataract-associated fibrosis through targeting STAT3

doi: 10.1016/j.gendis.2025.101549

Figure Lengend Snippet: The protein level of NR2F1 is increased in the ASC model. (A) The anesthetized and dilated mice were positioned beneath a microscope for the ASC modeling procedure. (B) Cataract lesions in control and ASC mice using a slit lamp. (C) Hematoxylin and eosin staining in lens section of control and ASC mice. Scale bar, 100 μm. (D) Masson staining of the two groups. Scale bar, 100 μm. (E) The immunofluorescence results of FN1 and VIM in lens slides of control and ASC groups. Scale bar, 50 μm. (F, G) The protein expression and quantification of apoptosis-related markers BAX and CASP3 in control and ASC groups. n = 3 per group; mean ± standard deviation; ∗∗ p < 0.01; unpaired student's t -test. (H, I) The protein level and quantitative chart of NR2F1 in the two groups mentioned above. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; unpaired student's t -test. NR2F1, nuclear receptor subfamily 2 group F member 1; ASC, anterior subcapsular cataract; FN1, fibronectin 1; VIM, vimentin; BAX, Bcl-2-associated X; CASP3, caspase 3.

Article Snippet: NR2F1 agonist 1 (HY-149913, MCE) was treated at 0.5 or 1 μM concentration.

Techniques: Microscopy, Control, Staining, Immunofluorescence, Expressing, Standard Deviation

Journal: Genes & Diseases

Article Title: Autophagy-induced NR2F1 activation promotes the apoptosis of lens epithelial cells and facilitates cataract-associated fibrosis through targeting STAT3

doi: 10.1016/j.gendis.2025.101549

Figure Lengend Snippet: TGF-β1 mediated autophagy dysfunction resulting in an increased protein level of NR2F1. (A) The mRNA profile of NR2F1 in lens epithelial cells and fiber cells using single-cell data. (B) The mRNA level of NR2F1 in SRA01/04 cells with PBS or TGF-β1. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; unpaired student's t -test. (C, D) The protein expression and quantification of NR2F1 in TGF-β1-induced SRA01/04 cells. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; unpaired student's t -test. (E) Subcellular location of NR2F1 in PBS- or TGF-β1-treated SRA01/04 cells. Scale bar, 50 μm. (F, G) The protein level and quantification of NR2F1 in SRA01/04 cells stimulated with the autophagy inhibitor chloroquine at concentrations of 5, 10, 20, 40, and 80 μM, respectively. n = 3 per group; mean ± standard deviation; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; one-way ANOVA. (H) Co-localization of LC3B and NR2F1 along with P62 and NR2F1 in SRA01/04 cells with TGF-β1. Scale bar, 50 μm. TGF-β1, transforming growth factor-β1; NR2F1, nuclear receptor subfamily 2 group F member 1; PBS, phosphate-buffered saline; LC3B, microtubule-associated protein 1 light-chain 3B.

Article Snippet: NR2F1 agonist 1 (HY-149913, MCE) was treated at 0.5 or 1 μM concentration.

Techniques: Standard Deviation, Expressing, Saline

Journal: Genes & Diseases

Article Title: Autophagy-induced NR2F1 activation promotes the apoptosis of lens epithelial cells and facilitates cataract-associated fibrosis through targeting STAT3

doi: 10.1016/j.gendis.2025.101549

Figure Lengend Snippet: Knockdown of NR2F1 significantly attenuates fibrosis both in vivo and in vitro . (A) Adeno-associated adenovirus (AAV) was administered into the anterior chamber of mouse eyes. (B) Representative pictures of the ASC modeling group versus the negative control group under a slit lamp following AAV injection. (C) Hematoxylin and eosin staining in the two groups mentioned above. Scale bar, 100 μm. (D) Masson staining in the two groups. Scale bar, 100 μm. (E) The immunofluorescence intensity of α-SMA in the lens sections of AAV-NC and AAV-NR2F1 ASC mice. Scale bar, 50 μm. (F, G) The protein level and quantification of NR2F1 in SRA01/04 cells transfected with Sh-NC, Sh-NR2F1-1, Sh-NR2F1-2, or Sh-NR2F1-3 lentivirus. n = 3 per group; mean ± standard deviation; ∗∗∗ p < 0.001; one-way ANOVA. (H–K) The protein expression and quantitative graphs of FN1, VIM, and α-SMA in TGF-β1-induced SRA01/04 cells with Sh-NC or Sh-NR2F1. n = 3 per group; mean ± standard deviation; ∗ p < 0.05, ∗∗ p < 0.01; unpaired student's t -test. (L – N) Immunofluorescence images of FN1, VIM, and α-SMA in TGF-β1-mediated SRA01/04 cells with Sh-NC or Sh-NR2F1. Scale bar, 100 μm. NR2F1, nuclear receptor subfamily 2 group F member 1; ASC, anterior subcapsular cataract; FN1, fibronectin 1; α-SMA, α-smooth muscle actin; VIM, vimentin; TGF-β1, transforming growth factor-β1.

Article Snippet: NR2F1 agonist 1 (HY-149913, MCE) was treated at 0.5 or 1 μM concentration.

Techniques: Knockdown, In Vivo, In Vitro, Negative Control, Injection, Staining, Immunofluorescence, Transfection, Standard Deviation, Expressing

Journal: Genes & Diseases

Article Title: Autophagy-induced NR2F1 activation promotes the apoptosis of lens epithelial cells and facilitates cataract-associated fibrosis through targeting STAT3

doi: 10.1016/j.gendis.2025.101549

Figure Lengend Snippet: NR2F1 inhibition suppresses epithelial cell apoptosis and migration. (A) The TUNEL staining in lens sections of AAV-NC and AAV-NR2F1 mice with ASC. Scale bar: 50 μm. (B – D) The protein levels and quantitative charts of BAX and CASP3 in the two groups mentioned above. n = 3 per group; mean ± standard deviation; ∗ p < 0.05, ∗∗ p < 0.01; unpaired student's t -test. (E, F) The TUNEL staining in the TGF-β1-mediated group with Sh-NR2F1 compared with that with Sh-NC. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; unpaired student's t -test; scale bar, 200 μm. (G – I) The protein expression and quantification of BAX and CASP3 in TGF-β1-induced SRA01/04 cells with or without Sh-NR2F1. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; unpaired student's t -test. (J, K) The transwell assay in the two groups mentioned above. n = 4 per group; mean ± standard deviation; ∗∗ p < 0.01; unpaired student's t -test. (I–K) The protein expression and quantification of BAX and CASP3 in TGF-β1-induced SRA01/04 cells with or without Sh-NR2F1. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; unpaired student's t -test. NR2F1, nuclear receptor subfamily 2 group F member 1; AAV, adeno-associated adenovirus; ASC, anterior subcapsular cataract; BAX, Bcl-2-associated X; CASP3, caspase 3; TGF-β1, transforming growth factor-β1.

Article Snippet: NR2F1 agonist 1 (HY-149913, MCE) was treated at 0.5 or 1 μM concentration.

Techniques: Inhibition, Migration, TUNEL Assay, Staining, Standard Deviation, Expressing, Transwell Assay

Journal: Genes & Diseases

Article Title: Autophagy-induced NR2F1 activation promotes the apoptosis of lens epithelial cells and facilitates cataract-associated fibrosis through targeting STAT3

doi: 10.1016/j.gendis.2025.101549

Figure Lengend Snippet: NR2F1 directly binds to STAT3 and regulates the expression of p-STAT3. (A, B) The protein levels and quantification of JAK1, p-STAT3, SMAD2, and MYD88 in TGF-β1-treated SRA01/04 cells with or without Sh-NR2F1. n = 3 per group; mean ± standard deviation; ns, >0.05; ∗∗ p < 0.01; unpaired student's t -test. (C, D) The protein level and quantitative chart of p-STAT3 in AAV-NC or AAV-NR2F1 ASC mice. n = 3 per group; mean ± standard deviation; ∗ p < 0.01; unpaired student's t -test. (E) The motif of NR2F1 predicted by the JASPAR website. (F) A dual-luciferase vector. (G) Mutant and wild-type STAT3 plasmids consequences. (H) The dual-luciferase assay for NR2F1 and STAT3 promoter. n = 3 per group; mean ± standard deviation; ns, >0.05; ∗ p < 0.05; unpaired student's t -test. (I, J) The protein level of p-STAT3 in TGF-β1-induced SRA01/04 cells following NR2F1 agonist treatment at 0.5 or 1 μM. n = 3 per group; mean ± standard deviation; ns > 0.05; ∗ p < 0.05; one-way ANOVA. (K) Structure of p-STAT3 specific inhibitor NSC 74859. (L, M) The protein expression and quantification of p-STAT3, STAT3, JAK1, FN1, VIM, and α-SMA in TGF-β1-induced SRA01/04 cells treated with the specific P-STAT3 inhibitor. n = 3 per group; mean ± standard deviation; ns, >0.05, ∗ p < 0.05, ∗∗ p < 0.01; one-way ANOVA. (N, O) The protein expression of the apoptosis-related markers BAX and CASP3 in TGF-β1-induced SRA01/04 cells treated with NSC 74859. n = 3 per group; mean ± standard deviation; ∗ p < 0.05; one-way ANOVA. NR2F1, nuclear receptor subfamily 2 group F member 1; STAT3, signal transducer and activator of transcription 3; p-STAT3, phosphorylated STAT3; ASC, anterior subcapsular cataract; FN1, fibronectin 1; α-SMA, α-smooth muscle actin; VIM, vimentin; BAX, Bcl-2-associated X; CASP3, caspase 3; TGF-β1, transforming growth factor-β1; JAK1, Janus kinase 1; SMAD2, SMAD family member 2; AAV, adeno-associated adenovirus.

Article Snippet: NR2F1 agonist 1 (HY-149913, MCE) was treated at 0.5 or 1 μM concentration.

Techniques: Expressing, Standard Deviation, Luciferase, Plasmid Preparation, Mutagenesis

Journal: Genes & Diseases

Article Title: Autophagy-induced NR2F1 activation promotes the apoptosis of lens epithelial cells and facilitates cataract-associated fibrosis through targeting STAT3

doi: 10.1016/j.gendis.2025.101549

Figure Lengend Snippet: The regulatory mechanism of NR2F1 in TGF-β1-induced SRA01/04 cells. NR2F1, nuclear receptor subfamily 2 group F member 1; TGF-β1, transforming growth factor-β1.

Article Snippet: NR2F1 agonist 1 (HY-149913, MCE) was treated at 0.5 or 1 μM concentration.

Techniques: