2D- and 3D-cultures of human trabecular meshwork cells: A preliminary assessment of an in vitro model for glaucoma study
Fig 4
Induction of pro inflammatory factors by chronic H2O2 treatment.
Quantitative PCR gene expression analysis of 2D- and 3D-HTMC subjected 500μM for 48 h and 72 h. IL-1α IL-1β, and IL-6 (Panels A, B and C, respectively). Data are expressed as fold-increase relative to the 2D control at the same end-point and normalized to Ubiquitin housekeeping gene expression. Each bar represents the mean ± S.D. of three independent experiments performed in triplicate. (Panel D) The figures depicted are representative of at least three similar immunoblot analysis of NF-kB (p65), p-NF-kB (p65) protein levels in untreated HTMCs and treated HTMCs (H2O2) whole protein lysates at indicated time points. GAPDH was used as an internal control for equal protein loading on the gel. (Panel E) NF-kBp65 activation was evaluated in HTMC cells subjected to chronic treatment with H2O2 for 24, 48 and 72 hrs. The analysis was performed by immunoblotting and the bars represent the ratio of phosfoNF-kBp65/NF-kBp65, and are expressed as fold vs. untreated HTMC cultures. Data represent the mean ± S.D. of 3 independent experiments. ***p <0.001,* p <0.05 treated 3D-HTMCs vs. untreated 3D-HTMCs cells; ### p<0.001, # p <0.05 untreated 3D-HTMC vs. untreated 2D-HTMC cells; §§§p<0.001 treated 3D-HTMC vs. treated 2D-HTMC cells (Two-way ANOVA followed by Bonferroni posttests).