Mir 30a

The isolated total RNA fraction was retro-transcribed using the Applied Biosystems™ High Capacity cDNA Reverse Transcription kit (Thermo Fisher Scientific, Waltham, MA, USA). Real-Time PCR was carried out using the Applied Biosystems® 7500 Real-Time PCR System. A list of primers is shown in Supplementary Table 6. The miRNAs were amplified individually using miR-10a (ID000387), miR-10b (ID002218), miR-143 (ID002249), miR-411 (ID001610), miR-181a (ID000480), miR-125a (ID002198), miR-125b-1* (ID002378), miR-127 (ID002229), miR-654 (ID002239), miR-30a (ID000417), let-7a (ID000377) and RNU44 (ID001094) assays (Invitrogen Life Technologies). Expressions were determined by C_T and expression fold change was calculated as previously described^{58 (link)}.

The lens capsular bags were cultured as previously described^{17 (link)}. Briefly, the capsular bags were put in DMEM-F12 with 10% fetal bovine serum containing 5 mM D-glucose (NG), 25 mM D-glucose (HG), or 5 mmol/L D-glucose plus 20 mmol/L mannitol (HM). After 3 days, they were transfected with a mimic, inhibitor or NC of miR-30a (Guangzhou RiboBio Co., Guangzhou, China) by using our previously reported method^{17 (link)}. In brief, the miR-30a mimic, inhibitor or NC was mixed with lipofectamineTM 2000 (Invitrogen, Carlsbad, CA) to form miRNA-lipofectamineTM 2000 complexes. Subsequently, the complexes were added in the medium at a final concentration of 50 nM and incubated with the capsular bags for 6 hours. They were also treated with both TGF-beta2) (10 ng/ml) and a miR-30a-5p mimic.