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. 2019 Nov 18;8(11):1455. doi: 10.3390/cells8111455

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Overexpression of miR-29b induces Bim mediated apoptosis. Cell lysate from (A) xenograft tumors treated with mimic miR-29b and (B) PC3 cells were analyzed by Western blot using specific antibody to Bim. Extra-large transcript of Bim (Bim_EL) was upregulated in mimic overexpressed samples. The membrane was reprobed with antibody to Actin as an internal control. Right panel shows quantitation of the data using Image J software. Densitometric scanning results are presented from three independent experiments. (C) PC3 cells were transfected with/without mimic miR-29b for 48 h and immune-stained with cytochrome C (red). DAPI (blue) was used for nuclearstaining. Representative confocal microscopic images showing increased cytoplasmic expression of cytochrome C in miR-29b transfected cells compared to control cells. Magnifications 60 X and scale bar 20 µm. Right panel shows quantitation of fluorescence intensity using ImageJ software using random five fields. Lysates of (D) xenograft tumors, (E) PC3 or (F) DU145 cells were subjected to Western blot analysis for PARP expression. Membrane was reprobed with Actin as internal control. Right panel shows quantitation of cleaved PARP using Image J software. Small bar indicates standard error (*, p < 0.05; *** p < 0.001).

Overexpression of miR-29b induces Bim mediated apoptosis. Cell lysate from (A) xenograft tumors treated with mimic miR-29b and (B) PC3 cells were analyzed by Western blot using specific antibody to Bim. Extra-large transcript of Bim (Bim_EL) was upregulated in mimic overexpressed samples. The membrane was reprobed with antibody to Actin as an internal control. Right panel shows quantitation of the data using Image J software. Densitometric scanning results are presented from three independent experiments. (C) PC3 cells were transfected with/without mimic miR-29b for 48 h and immune-stained with cytochrome C (red). DAPI (blue) was used for nuclearstaining. Representative confocal microscopic images showing increased cytoplasmic expression of cytochrome C in miR-29b transfected cells compared to control cells. Magnifications 60 X and scale bar 20 µm. Right panel shows quantitation of fluorescence intensity using ImageJ software using random five fields. Lysates of (D) xenograft tumors, (E) PC3 or (F) DU145 cells were subjected to Western blot analysis for PARP expression. Membrane was reprobed with Actin as internal control. Right panel shows quantitation of cleaved PARP using Image J software. Small bar indicates standard error (*, p < 0.05; *** p < 0.001).