Research Paper Volume 11, Issue 9 pp 2699—2723

Laminin α4 overexpression in the anterior lens capsule may contribute to the senescence of human lens epithelial cells in age-related cataract

Figure 5. MMP-9 reduces cell senescence and laminin (LM) deposition induced by H2O2. Senescent HLE B-3 cells [or cell basement membranes (BMs)] were cultured in medium with H2O2 (400 μM) for 96 h. (A-I) HLE B-3 cells were treated with H2O2 only, or in combination with indicated plasmid. (A) Immunoblot analysis of MMP-9 in HLE B-3 cells. (B-C) Percentage of SA-β-gal-positive cells (B) and protein expression of GLB1 (C) in HLE B-3 cells. (D) Total LM in HLE B-3 cells, detected by ELISA. (E) Protein expressions of TGF-β1 and LM subunits in HLE B-3 cells, detected by IB. (F) Total LM in HLE B-3 cell BMs, detected by ELISA. (G) SDS-PAGE analysis followed by CBB staining of HLE B-3 cell BMs. (H) Immunoblot analysis of LMα4 and LMα1 in HLE B-3 cell BMs. (I) Immunofluorescence analysis of LMα4 (green) and LMβ3 (green) in HLE B-3 cell BMs (Scale bars: 50 μm). (J-M) HLE B-3 cells were treated with H2O2 only, or in combination with indicated siRNA. (J) The percentage of SA-β-gal-positive cells in HLE B-3 cells. (K) Total LM in HLE B-3 cells, as detected by ELISA. (L) Immunoblot analysis of LM subunits and TGF-β1 in HLE B-3 cells. (M) Total LM in HLE B-3 cell BMs, as detected by ELISA. Data are shown as mean ± SD and were analyzed using paired t-test. *, p<0.05; ***, p<0.001.