Figure 3. Relative level of DC6 cDNAs expression after six days of cytokine treatment, in RCS rat retina and in Müller glial cell cultures

A: Retinal explants from P0 rat were cultured with medium alone (white bars) or with 10 ng/ml of either CNTF (red bars) or LIF (black bars). Retinal explants were harvested after 6DIV and RNA were extracted. cDNA expression corresponding to 13 clones (F5, G4, E2, D2, A7, F2, D1, C2, B6, B2, G7, C7, and A2) isolated from DC6 library was analyzed by real-time RT-PCR experiments. For each gene, all values were compared to mRNA expression of untreated rat retinal explants at 6DIV. Data are expressed as mean±SD from two independent experiments performed in triplicate. (***p<0.001; **p<0.01; *p<0.05) B: Retinas from control RCS rat (white bars) and dystrophic RCS rat (black bars) were collected at three months and RNA extracted. cDNA expression corresponding to rhodopsin and to nine clones (A7, F2, D1, C2, B6, B2, G7, C7, and A2) isolated from the DC6 library and differentially expressed upon addition of CNTF or LIF, was analyzed by real-time PCR. For each gene, all values were compared to mRNA expression of control RCS rat retina. Data are expressed as mean±SD from two independent experiments performed in triplicate. (***p<0.001; **p<0.01; *p<0.05). C: RNA was extracted from pure retinal Müller cell culture and cDNA expression corresponding to nine clones (A7, F2, D1, C2, B6, B2, G7, C7, and A2) analyzed by real-time PCR. The expression of vimentin (Vim), a glial specific gene, was assessed as a control and arbitrarily fixed at 10,000 u.a. The cDNAs from DC6 library were classified in three groups based on their level of transcription ranged from 6 to 1255 u.a.: Low (white bars, 500 fold less transcripts detected than vimentin transcripts), Intermediate (Int; red bars, 50 fold) and high (black bars, 5 fold).