Samples of conjunctival (including Tenon's capsule) and sclera tissues were collected from the five groups. These samples were washed thrice using ice-cold PBS. Then, they were extracted in cold RIPA lysis buffer (strong), which consisted of 20 mmol/L Tris (pH 7.5), 150 mmol/L NaCl, 1% Triton X-100, 2.5 mmol/L sodium pyrophosphate, 1 mmol/L EDTA, 1% Na3VO4, 5 μg/mL leupeptin, and 1 mmol/L phenyl methyl sulfonyl fluoride (PMSF). After centrifugation for 10 minutes at 16099g, we collected the supernatant and used it in Western blot analysis. We visualized the immunoreactive proteins on autoradiograph films using chemiluminescence detection reagents (ECL; GE Healthcare, Laurel, MD, USA). Monoclonal antibodies to PI3Kp85α, phospho-PI3Kp85α, Akt, phospho-Akt, Sp1, and Col1A1 were obtained commercially (Cell Signaling Technologies, Danvers, MA, USA; GE Healthcare). The antibody β-Actin (Sigma-Aldrich Corp., St. Louis, MO, USA) was used as the loading control in all cases.