Fig 1: Immunohistochemical staining for ABC transporters at the CSF/brain ventricular interface in the rat. This layer changes from a neuroepithelium in the developing brain to ependyma in the adult. Coronal sections through lateral ventricles of postnatal P4 (a), P10-14 (c,e) and adult cortex (b,d,f). Sections are immunostained with antibodies to: MRP5 (abcc5) for (a,b), PGP (abcb1a/b) for (c,d) or BCRP (abcg2) for (e,f). Dorsal side is up. Note distinct immunostaining, indicated by filled arrowheads, of MRP5 in the adult brain (b) and PGP/BCRP in the postnatal brain (c,e). This is in contrast to the absence of immunostaining, indicated by unfilled arrowheads, of PGP and BCRP in the adult (d,f). MRP5 staining in the postnatal brain (a) is lightly present in some regions, filled arrowheads, and absent in others, unfilled arrowhead. Also note positive immunostaining in choroid plexuses in (b,e,f) (indicated by *). For age-related differences in the immunostaining for other ABC-transporters see Table 1. Scale bar 100 µm (bottom right) applies to all images.
Fig 2: Inhibition of FOXM1 suppresses colony formation and induces apoptosis through downregulation of ABCC5/10. (A) mRNA and (B) protein levels of FOXM1 were downregulated by siRNA knockdown. (C) Knockdown of FOXM1 resulted in impaired colony formation (D) Knockdown of FOXM1 in resistant cells increased caspase 3/7 activity (n=3; **P<0.01). (E) Protein levels of ABCC5/10 in parental, resistant cells, miR-361-overexpressing and FOXM1 knockdown cells. (F) Reduced expression of miR-361 in colorectal cancer cells contributes to 5-FU chemoresistance by activating the FOXM1-ABCC5/10 signaling pathway. FOXM1, forkhead box M1; ABCC, ATP binding cassette subfamily C; si(RNA), small interfering (RNA); NC, negative control; miR, microRNA; 5-FU, 5-fluorouracil; Res, resistant.
Fig 3: Cocaine Modulates Drug Transporter Expression.Immunofluorescent microscopy was performed to evaluate (A) BCRP, (B) OAT1, (E) ENT1, (F) OAT3, (I) MRP1, (J) OATP1A2, (M) MRP4, (N) OATP2A1, (Q) MRP5, or (R) P-gp (green) following treatment with cocaine (10 µM, right) or vehicle (left) for 24 hours. DAPI was used to visualize nucleus (blue). One paired representative image, out of 20 individual images, are shown. All scale bars = 50 µm. (C, D, G, H, K, L, O, P, S, T) Quantification of the fluorescent signal from immunofluorescent microscopy was performed for endothelial cells treated with cocaine (10 µM, burgundy) or vehicle (teal) for 24 hours. Twenty independent experiments (represented by individual dots) were performed. Estimation plots are shown where the left y-axis denotes relative fluorescent intensity (RFU, pixels) and the right y-axis reflects the effect size (black bar), which is the difference between means of each condition. Data are represented as mean ± standard deviation. *p<0.05. ***p<0.001. ****p<0.0001. Unpaired T-test was performed.
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