Figure 1.

PKA activation protects lens fiber cells against H₂O₂-induced cell death via functional Cx50 HCs and GSH transport

(A–C) Primary chick lens cell cultures were infected with high-titer recombinant RCAS(A) retroviruses containing Cx50, or Cx50 mutants H156N, and then treated with or without PKA activator: forskolin (1 μM) for 2 h before being subjected to fluid flow shear stress (FFSS) at 1 dyne/cm² for 30 min, incubated with 1 mM GSH for 10 min and treated with 50 μM H₂O₂ for 4 h. Cell apoptosis and necrosis were detected using Dead Cell Apoptosis Kit. The percentage of cells under apoptosis (FITC-Annexin V+) (B) and PI + cells (C) was quantified (n = 4). Scale bar: 50 μm ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 (two-way ANOVA). At least three microphotographs of fluorescence fields were captured by a 20X microscope (Keyence BZ-X710) with a FITC filter and a rhodamine filter.