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. Author manuscript; available in PMC: 2013 Jul 29.
Published in final edited form as: J Neurochem. 2009 Aug 3;111(1):250–263. doi: 10.1111/j.1471-4159.2009.06322.x

Fig. 1.

Fig. 1

(a) CRX and OTX2 expression in retinoblastoma cell lines. Western blot analysis of retinoblastoma cell lines using anti-CRX and anti-OTX2 antibodies. Fifty μg of whole cell lysates were loaded in each lane and electrophoresed in a 12% polyacrylamide-SDS gel. Proteins were transferred to PVDF membranes and sequentially immunostained with rabbit anti-CRX, followed by rabbit anti-OTX2 antibody (after stripping the blot), and goat anti-actin antibody. The signal was detected using the ECL or Immobilon detection reagents. (b) Northern blot analysis of CRX and OTX2 in retinoblastoma cell lines. Poly(A)+ RNA was isolated from 12 retinoblastoma cell lines. Two μg poly(A)+ RNA were loaded in each lane. The RNA was electrophoresed in an agarose-formaldehyde gel and the RNA transferred to a nitrocellulose filter. The filter was sequentially hybridized with 32P-labeled CRX, OTX2 and β-actin cDNAs. Sizes of the transcripts are indicated on the side. (c) Northern blot analysis of CRX/OTX2 target genes in retinoblastoma cell lines. Two μg poly(A)+ RNA from each of the indicated retinoblastoma cell lines and calf retina were electrophoresed in a 1.5% agarose-formaldehyde gel and transferred to a nitrocellulose filter. The filter was sequentially hybridized with 32P-labeled IRBP, PDEB, arrestin1, cone arrestin and β-actin cDNAs. The latter served as the loading control. Sizes of the transcripts are indicated on the side.