1：Experimental Design and Method Selection:

The study used in vitro experiments with human lens epithelium (HLE)-B3 cells treated with TGF-β2 and Y-27632 to assess mRNA expression, and in vivo experiments with mice exposed to UVR-B to induce cataracts and treated topically with Y-27632 to evaluate cataract formation.

2：Sample Selection and Data Sources:

HLE-B3 cells were purchased from the American Type Culture Collection. Female C57BL/6Cr Slc mice (12 weeks old) were used.

3：List of Experimental Equipment and Materials:

Equipment includes transilluminator (TFML-20; UVP), radiometer (UV-340; Custom), slit lamp microscope (SL-15; Kowa), microscope and camera system, Eco RT-PCR system (Illumina). Materials include DMEM (Nacalai Tesque), FBS (GE Healthcare Life Sciences), TGF-β2 (Sigma-Aldrich), Y-27632, ISOGEN reagent (Nippon Gene), PrimeScript Reverse Transcriptase (Takara Bio), Ex Taq polymerase (Takara Bio), antibodies (anti-α-SMA from Dako, anti-type I collagen from Abcam), Vectastain ABC Standard kit (Vector Laboratories).

4：Experimental Procedures and Operational Workflow:

For in vitro, HLE-B3 cells were cultured and treated with TGF-β2 and Y-27632 for 24 hours, then RNA was extracted for qRT-PCR. For in vivo, mice were exposed to UVR-B twice a week for 3 weeks, with Y-27632 or saline instilled topically twice daily starting 3 days before exposure. Lens opacity was evaluated with a slit lamp and microscope, and lenses were processed for histopathology and immunohistochemistry.

5：Data Analysis Methods:

Data were analyzed using unpaired Student's t-test or one-way ANOVA with Tukey's test, using Kaleida Graph software v.4.1.