Skip to main content
Figure 4 | Molecular Cancer

Figure 4

From: RNA-binding proteins to assess gene expression states of co-cultivated cells in response to tumor cells

Figure 4

RNase Protection Assay (RPA) of mRNAs from mixed mouse and human cell lines. Species specificity of multiprobe RPAs using: A) the human tumor suppressor probe set (hTS1), and B) the mouse angiogenesis RPA probe set (mAngio), was verified using total RNA extracted from murine PY4.1 cells and human T98G cells. Unprotected RNA probes were used to identify the nature of the different sized protected fragments. C) RPA gene expression profile of PY4.1 cells expressing G10-PABP obtained with total RNA and with mRNA derived from immunoprecipitations with anti-PABP serum or anti-G10 antibody D) RPA gene expression profile of T98G cells expressing Flag-PABP obtained from total RNA and from mRNA derived from immunoprecipitations with anti-PABP serum or anti-Flag antibody. E, F and G) Mixed extracts from cells expressing T98G Flag-PABP and PY4.1 G10-PABP were immunoprecipitated with anti-PABP serum or anti-Flag or anti-G10 antibodies. The mRNA populations generated by immunoprecipitations were analyzed with RPA of both the mouse angiogenesis (mAngio) and the human tumor suppressor probe (hTS1) sets. GAPDH and L32 are controls in both probe sets and show cross species hybridization. The asterisk in B and G also indicate a band resulting from cross species hybridization. The experiments indicate that species-specific mRNA populations can be isolated and quantified by the use of distinct tagged-PABPs.

Back to article page