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Figure 2 | Journal of Molecular Signaling

Figure 2

From: Direct RNA sequencing mediated identification of mRNA localized in protrusions of human MDA-MB-231 metastatic breast cancer cells

Figure 2

RNA in MDA- MB- 231 protrusions. (A-B) Ethidium bromide (EtBr) mediated visualization of RNA in protrusions of MDA-MB-231 cells grown in Boyden chamber. (A) RNase treated cells (+ RNase) are shown for control of RNA specific EtBr staining. The left panels show DAPI staining, central pictures EtBr staining and merged pictures are shown in the right panels. Arrowheads exemplify EtBr staining in cell protrusions. Scale bar 20 μm. (B) EtBr mediated visualization of RNA localization in protrusions migrated through the 1 μm pore size membrane in the Boyden chamber. The cell bodies on the upper side of the membrane seen in (A) were removed by wiping and DAPI staining was performed in parallel to the pictures shown in (A) (left panel). EtBr staining was used to visualize cell material at the membrane lower side (arrowheads, right panel). Exposure times were increased compared to (A) visualizing also the 1 μm membrane pores. Scale bar 20 μm. (C) Determination of the relative RNA localization ratio in MDA-MB-231 protrusions by RT-qPCR analyses of candidate genes. The RNA localization ratio was calculated as the relative expression level in the protrusions fraction compared to the cell body fraction. The localization ratio of RNA in protrusions was normalized to the RNA localization ratio of ARPC3 given the value 1.

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Journal of Molecular Signaling

ISSN: 1750-2187