Figure 5.

Bcl-2 mRNA expression of ONHA and TMC after treatment with minocycline 20 μM and 40 μM only or after additional treatment with 600 mM H₂O₂ or TGFβ-2, as investigated by quantitative RT-PCR. All tests were performed in triplicate and repeated three times. The mRNA levels were normalized to those of 18S rRNA and expressed as the ratio of Bcl-2 mRNA/18S rRNA. ONHA treated with minocycline concentrations showed a significant increase in Bcl-2 mRNA expression compared with the control in both cell lines. TGFβ-2 and H₂O₂ treatment decreased Bcl-2 expression. When ONHA cells were treated with minocycline 20 μM and 40 μM and TGFβ-2 or H₂O₂, Bcl-2 expression was increased compared with ONHA treated only with TGFβ-2 or H₂O₂. Similarly, TMC treated with minocycline 20 μM and TGFβ-2 or H₂O₂ showed an increased expression of Bcl-2 mRNA compared with TMC treated only with TGFβ-2 or H₂O₂. All differences between the two minocycline concentrations and the corresponding controls are statistically significant.

Key: x-axis, RR of Bcl-2 mRNA normalized to 18s rRNA expressed in decimal format; y-axis, tested concentrations of minocycline.

Abbreviations: TMC, trabecular meshwork cells; ONHA; optic nerve head astrocytes; RT-PCR, real time polymerase chain reaction; RR, relative ratio; TGFβ-2, transforming growth factor-beta-2.