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. 2015 Jun 26;7(7):3443–3461. doi: 10.3390/v7072780

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© 2015 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 license (http://creativecommons.org/licenses/by/4.0/).

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E7 does not co-localise with an autophagosome marker, LC3 (A) but appears to co-localise with endoplasmic reticulum marker calreticulin (B) in the presence of A2. SiHa cells were mock-transfected (M) or transfected with 100 nM A2 or SF1 using Oligofectamine. At 14 h post-transfection, cells were co-stained with anti-E7 (Abcam) and either anti-LC3 (an autophagosomal marker) or anti-calreticulin (an ER marker) prior to incubation with either AlexaFluor568 rabbit anti-mouse or AlexaFluor488 goat anti-rabbit secondary antibodies. Red, Green and Blue are E7, FITC (LC3, autophagosome) and DAPI (nucleus), respectively. The last panel is a zoom of merged images. The scale bar = 10 µm. E7/LC3 and E7/calreticulin colocalisation was analysed using Bitplane:Imeris image analysis software based on the calculation of Pearson’s correlation coefficient values and presented as percentage co-localisation (C). n = 21, 23 and 17 for E7/LC3 and n = 19, 39 and 20 for E7/calreticulin with mock-, A2- and SFI-treatment, respectively. * p ≤ 0.0005, ** p ≤ 0.002.

E7 does not co-localise with an autophagosome marker, LC3 (A) but appears to co-localise with endoplasmic reticulum marker calreticulin (B) in the presence of A2. SiHa cells were mock-transfected (M) or transfected with 100 nM A2 or SF1 using Oligofectamine. At 14 h post-transfection, cells were co-stained with anti-E7 (Abcam) and either anti-LC3 (an autophagosomal marker) or anti-calreticulin (an ER marker) prior to incubation with either AlexaFluor568 rabbit anti-mouse or AlexaFluor488 goat anti-rabbit secondary antibodies. Red, Green and Blue are E7, FITC (LC3, autophagosome) and DAPI (nucleus), respectively. The last panel is a zoom of merged images. The scale bar = 10 µm. E7/LC3 and E7/calreticulin colocalisation was analysed using Bitplane:Imeris image analysis software based on the calculation of Pearson’s correlation coefficient values and presented as percentage co-localisation (C). n = 21, 23 and 17 for E7/LC3 and n = 19, 39 and 20 for E7/calreticulin with mock-, A2- and SFI-treatment, respectively. * p ≤ 0.0005, ** p ≤ 0.002.