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. 2014 Mar 5;9(3):e89772. doi: 10.1371/journal.pone.0089772

Figure 5. Rosemary extract modulates ER stress proteins.

Figure 5

A) Cell lysates of 22Rv1 and LNCaP cells were prepared after treatment with RE-40 for 24hrs. Cell lysates were used to evaluate expression of ER stress proteins PERK, IRE1α, BiP and CHOP using Western blotting. Equal loading of protein was confirmed by probing the blot with β-actin. B) Splicing of XBP1 mRNA as an indicator of ER stress activation in both 22Rv1 and LNCaP treated cells. GAPDH was used as an internal control for RT-PCR. C) 22Rv1 and LNCaP cell lysates were tested for caspase-4 activation during ER stress by western blotting. D) Immunofluorescence staining for CHOP expression in 22Rv1 and LNCaP prostate cancer cells after treatment with 50 µg/ml of RE-40. For both cell lines the first column shows: DAPI-stained nuclei appear blue, second column: Tx-red stained nuclei appear red displaying expression of CHOP protein, third column: merged picture of DAPI and Tx-red. E) To determine the effect of RE-40 on human cells, prostate epithelial cells were treated with RE-40 and lysates confirmed for expression of ER stress protein, PERK and CHOP, by Western blotting