Fig. 2
From: Epigenetic silencing of a long non-coding RNA KIAA0495 in multiple myeloma
Methylation and expression of KIAA0495 in myeloma cells. a Semi-quantitative RT-PCR analysis showed expression of KIAA0495 in MOLP-8, RPMI-8226, U-266, WL-2, and JJN-3, which were completely unmethylated for KIAA0495, but not in KMS-12-PE, LP-1, NCI-H929, OCI-MY5, and OPM-2, which were partially methylated for KIAA0495 (P = 0.0079). Sequencing analysis confirmed the authenticity of the RT-PCR of KIAA0495. b qRT-PCR analysis showed the expression of KIAA0495 was significantly higher, as indicated by a lower ΔCt, in methylated than completely unmethylated myeloma cell lines (P = 0.000155). c Two partially methylated myeloma cell lines, LP-1 and OCI-MY5, were separately treated with 1 μmol/l of 5-AzadC for 3 days, after which, 5-AzadC was removed and cells were cultured for additional 6 days. On day 3, upon treatment with 5-AzadC, KIAA0495 was demethylated as indicated by the decreased percentage of methylation intensity by quantitative bisulfite pyrosequencing, with concommitent KIAA0495 re-expression as measured by qRT-PCR. On day 9, upon removal of 5-AzadC for 6 days, methylation of KIAA0495 was restored with re-suppression of KIAA0495