Journal: Oncology Reports

Article Title: Quercetin reduces expression of ATP-binding cassette transporters by regulating the PTEN/PI3K/AKT signaling pathway in breast cancer cells

doi: 10.3892/or.2026.9068

Figure Lengend Snippet: Effects of Que on cell viability, mRNA levels, apoptosis and protein expression in MCF-7 cells. (A) Cell viability of MCF-7 cells treated with the indicated concentrations of Que for 24 and 48 h, as determined by the Cell Cycle Kit-8 assay. (B) mRNA expression levels of ABCB1, ABCC2 and ABCG2 following Que treatment. (C) Representative western blotting images, (D) protein expression levels of ABCB1, ABCC2 and ABCG2, and (E) Bax, Bcl-2, and cleaved caspase-3 were assessed by western blotting. (F) Representative images of flow cytometry and (G) apoptosis rate was analyzed by flow cytometry after Annexin V-FITC/PI staining. Data are presented as the mean ± SEM of at least three independent experiments. Statistical significance was determined by one/two-way ANOVA followed by Bonferroni's post hoc test for multiple comparisons. *P<0.05, **P<0.01 compared with the control group, ## P<0.01 compared with the Que (24 h) group. Que, Quercetin; Ctrl, control.

Article Snippet: Primary antibodies included PTEN (1:5,000; cat. no. ab267787), Bcl-2 (1:6,000; cat. no. ab196495), Bax (1:6,000; cat. no. ab32503) and ABCG2 (1:5,000; cat. no. ab108312) from Abcam; p-PI3K (p85) (1:2,000; cat. no. 4228), p-AKT (Ser473) (1:2,000; cat. no. 4060), cleaved caspase-3 (1:5,000; cat. no. 9664), ABCC2 (1:1,000; cat. no. 12559) and ABCB1 (1:2,000; cat. no. 13342) from Cell Signaling Technology, Inc.; and GAPDH (1:12,000; cat. no. 60004-1-Ig), PI3K (1:3,000; cat. no. 20584-1-AP), AKT (1:3,000; 10176-2-AP), Caspase-3 (1:3,000; 19677-1-AP) and β-actin (1:10,000; cat. no. 66009-1-Ig) from Proteintech Group, Inc. After incubation with HRP-conjugated secondary antibodies (1:10,000; cat. no. SA00001-1 and SA00001-2; Proteintech Group, Inc.), protein bands were visualized using an ECL reagent (Beijing Lanbolide Trading Co., Ltd.) and imaged with a ChemiDoc XRS+ system (Bio-Rad Laboratories, Inc.).

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Control

Journal: Oncology Reports

Article Title: Quercetin reduces expression of ATP-binding cassette transporters by regulating the PTEN/PI3K/AKT signaling pathway in breast cancer cells

doi: 10.3892/or.2026.9068

Figure Lengend Snippet: Effects of PTEN overexpression in MCF-7 cells. mRNA expression levels of (A) PTEN, (B) ABCB1, ABCC2, ABCG2, Bax and Bcl-2 following PTEN overexpression. (C) Representative western blotting images and protein expression levels of (D) ABCB1, ABCC2, ABCG2, (E) PTEN, Bax and Bcl-2 following PTEN overexpression. The results are presented as the mean ± SEM of at least three independent experiments and analyzed using one-way ANOVA followed by Bonferroni's post hoc test. *P<0.05, **P<0.01 compared with the Plvx-con group.

Article Snippet: Primary antibodies included PTEN (1:5,000; cat. no. ab267787), Bcl-2 (1:6,000; cat. no. ab196495), Bax (1:6,000; cat. no. ab32503) and ABCG2 (1:5,000; cat. no. ab108312) from Abcam; p-PI3K (p85) (1:2,000; cat. no. 4228), p-AKT (Ser473) (1:2,000; cat. no. 4060), cleaved caspase-3 (1:5,000; cat. no. 9664), ABCC2 (1:1,000; cat. no. 12559) and ABCB1 (1:2,000; cat. no. 13342) from Cell Signaling Technology, Inc.; and GAPDH (1:12,000; cat. no. 60004-1-Ig), PI3K (1:3,000; cat. no. 20584-1-AP), AKT (1:3,000; 10176-2-AP), Caspase-3 (1:3,000; 19677-1-AP) and β-actin (1:10,000; cat. no. 66009-1-Ig) from Proteintech Group, Inc. After incubation with HRP-conjugated secondary antibodies (1:10,000; cat. no. SA00001-1 and SA00001-2; Proteintech Group, Inc.), protein bands were visualized using an ECL reagent (Beijing Lanbolide Trading Co., Ltd.) and imaged with a ChemiDoc XRS+ system (Bio-Rad Laboratories, Inc.).

Techniques: Over Expression, Expressing, Western Blot

Journal: Oncology Reports

Article Title: Quercetin reduces expression of ATP-binding cassette transporters by regulating the PTEN/PI3K/AKT signaling pathway in breast cancer cells

doi: 10.3892/or.2026.9068

Figure Lengend Snippet: Effects of combined PTEN overexpression and Que treatment on ABC transporters and the PI3K/AKT signaling pathway in MCF-7 cells. mRNA expression levels of (A) ABCB1, (B) ABCC2 (C) and ABCG2 in different treatment groups. (D) Representative images and (E) protein expression levels of ABCB1, ABCC2, ABCG2, (F) p-PI3K, and (G) p-AKT were evaluated by western blotting. The results are presented as the mean ± SEM of at least three independent experiments and analyzed using one-way ANOVA followed by Bonferroni's post hoc test. *P<0.05, **P<0.01, ***P<0.001 compared with the Plvx-con group. Que, Quercetin; con, control; p, phosphorylated.

Article Snippet: Primary antibodies included PTEN (1:5,000; cat. no. ab267787), Bcl-2 (1:6,000; cat. no. ab196495), Bax (1:6,000; cat. no. ab32503) and ABCG2 (1:5,000; cat. no. ab108312) from Abcam; p-PI3K (p85) (1:2,000; cat. no. 4228), p-AKT (Ser473) (1:2,000; cat. no. 4060), cleaved caspase-3 (1:5,000; cat. no. 9664), ABCC2 (1:1,000; cat. no. 12559) and ABCB1 (1:2,000; cat. no. 13342) from Cell Signaling Technology, Inc.; and GAPDH (1:12,000; cat. no. 60004-1-Ig), PI3K (1:3,000; cat. no. 20584-1-AP), AKT (1:3,000; 10176-2-AP), Caspase-3 (1:3,000; 19677-1-AP) and β-actin (1:10,000; cat. no. 66009-1-Ig) from Proteintech Group, Inc. After incubation with HRP-conjugated secondary antibodies (1:10,000; cat. no. SA00001-1 and SA00001-2; Proteintech Group, Inc.), protein bands were visualized using an ECL reagent (Beijing Lanbolide Trading Co., Ltd.) and imaged with a ChemiDoc XRS+ system (Bio-Rad Laboratories, Inc.).

Techniques: Over Expression, Expressing, Western Blot, Control

Journal: Oncology Reports

Article Title: Quercetin reduces expression of ATP-binding cassette transporters by regulating the PTEN/PI3K/AKT signaling pathway in breast cancer cells

doi: 10.3892/or.2026.9068

Figure Lengend Snippet: Effects of combined LY294002 and Que treatment on apoptosis and protein expression in MCF-7 cells. (A) Representative western blotting images and Protein expression levels of (B) ABCB1, ABCC2, ABCG2, (C) p-PI3K (D) PTEN, Bax, Bcl-2 and (E) p-AKT in different treatment groups. (F) mRNA expression levels of ABCB1, ABCC2, ABCG2, PTEN, Bax and Bcl-2 in different treatment groups. (G) Representative immunofluorescence images of ABCG2 expression (green). Nuclei were counterstained with DAPI (blue). Scale bar, 50 µm. (H) Representative flow cytometry images and (I) Apoptosis rate was analyzed by flow cytometry after Annexin V-FITC/PI staining. The results are presented as the mean ± SEM of at least three independent experiments and analyzed using two-way ANOVA followed by Bonferroni's post hoc test. *P<0.05, **P<0.01 compared with the control group; ## P<0.05 compared with the Que group. Que, Quercetin; Ctrl, control; p, phosphorylated.

Article Snippet: Primary antibodies included PTEN (1:5,000; cat. no. ab267787), Bcl-2 (1:6,000; cat. no. ab196495), Bax (1:6,000; cat. no. ab32503) and ABCG2 (1:5,000; cat. no. ab108312) from Abcam; p-PI3K (p85) (1:2,000; cat. no. 4228), p-AKT (Ser473) (1:2,000; cat. no. 4060), cleaved caspase-3 (1:5,000; cat. no. 9664), ABCC2 (1:1,000; cat. no. 12559) and ABCB1 (1:2,000; cat. no. 13342) from Cell Signaling Technology, Inc.; and GAPDH (1:12,000; cat. no. 60004-1-Ig), PI3K (1:3,000; cat. no. 20584-1-AP), AKT (1:3,000; 10176-2-AP), Caspase-3 (1:3,000; 19677-1-AP) and β-actin (1:10,000; cat. no. 66009-1-Ig) from Proteintech Group, Inc. After incubation with HRP-conjugated secondary antibodies (1:10,000; cat. no. SA00001-1 and SA00001-2; Proteintech Group, Inc.), protein bands were visualized using an ECL reagent (Beijing Lanbolide Trading Co., Ltd.) and imaged with a ChemiDoc XRS+ system (Bio-Rad Laboratories, Inc.).

Techniques: Expressing, Western Blot, Immunofluorescence, Flow Cytometry, Staining, Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Grouping cells uptake and efflux substrate experiments for each transporter protein in HEK-293 cells.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Primer sequences of OCT2, OAT1, OAT3, P-gp, MATE1, MATE2K, MRP2 genes.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Values of the binding energies of berberine and berberrubine to seven transporter proteins in the kidney (Kcal/mol).

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Binding Assay

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Molecular docking illustrations of BBR and BBRR with seven transport proteins respectively. (A) BBR docked with OAT1. (B) BBR docked with OAT3. (C) BBR docked with OCT2. (D) BBR docked with MATE1. (E) BBR docked with MATE2K. (F) BBR docked with P‐gp. (G) BBR docked with MRP2. (H) BBRR docked with OAT1. (I) BBRR docked with OAT3. (J) BBRR docked with OCT2. (K) BBRR docked with MATE1. (L) BBRR docked with MATE2K. (M) BBRR docked with P‐gp. (N) BBRR docked with MRP2.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Trends of metformin, methotrexate, estrone-3-sulfate, topotecan, rhodamine 123, and vinblastine concentration under different dosing time conditions in cells lysates or extracellular medium from HEK-293 cells. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR). (A) The trend of MET concentration changes mediated by OCT2. (B) The trend of MTX concentration changes mediated by OAT1. (C) The trend of E3S concentration changes mediated by OAT3. (D) The trend of Rho123 concentration changes mediated by P-gp. (E) The trend of TOP concentration changes mediated by MATE1 and MATE2K. (F) The trend of VBL concentration changes mediated by MRP2 (n = 3).

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Concentration Assay, Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: The effect of various solutions on each substrate accumulation transported by renal transport proteins. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR) (A) The effect of various solutions on the accumulation of MET transported by OCT2. (B) The effect of various solutions on the accumulation of MTX transported by OAT1. (C) The effect of various solutions on the accumulation of E3S transported by OAT3. (D) The effect of various solutions on the accumulation of Rho123 transported by P-gp. (E) The effect of various solutions on the accumulation of TOP transported by MATE1 and MATE2K. (F) The effect of various solutions on the accumulation of VBL transported by MRP2. Compared with the RPC group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; compared with the SPC group, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 (n = 3).

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: The effect of various solutions on the protein expression of MATE1, MATE2K, P-gp, MRP2. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR). (A) MATE1, MATE2K, P-gp, MRP2 protein expression bands in HEK-293 cells. (B) The effect of various solutions on MATE1 protein expression level. (C) The effect of various solutions on MATE2K protein expression level. (D) The effect of various solutions on P-gp protein expression level. (E) The effect of various solutions on MRP2 protein expression level. Compared with the BC group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; compared with the RPC group, □ p < 0.05, □□ p < 0.01, □□□ p < 0.001, □□□□ p < 0.0001; compared with the BBR group, △ p < 0.05, △△ p < 0.01, △△△ p < 0.001, △△△△ p < 0.0001.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: The effect of various solutions to MATE1, MATE2K, P-gp, MRP2 mRNA expression level. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR). (A) The effect of various solutions on MATE1 mRNA expression level. (B) The effect of various solutions on MATE2K mRNA expression level. (C) The effect of various solutions on P-gp mRNA expression level. (D) The effect of various solutions on MRP2 mRNA expression level Compared with the BC group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; compared with the RPC group, □ p < 0.05, □□ p < 0.01, □□□ p < 0.001, □□□□ p < 0.0001; compared with the BBR group, △ p < 0.05, △△ p < 0.01, △△△ p < 0.001, △△△△ p < 0.0001.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: mBio

Article Title: Guanylate-binding protein 5 antagonizes viral glycoproteins independently of furin processing

doi: 10.1128/mbio.02086-24

Figure Lengend Snippet: GBP5 expression increases the electrophoretic mobility of cell-associated HIV-1 Env, MLV Env, and SARS-CoV-2 S and reduces their incorporation into virions. HEK293T cells were co-transfected with varying quantities of a GBP5 expression vector, with total DNA held constant (at 0.5 µg) with empty vector control and ( A ) the pNL4-3 molecular clone (4 µg), ( B ) the Env-defective pNL4-3 derivative pNL4-3/KFS (3.3 µg) and MLV Env expression plasmid (0.6 µg), or ( C ) the Env(−) luciferase-encoding NL4-3-derived vector pNL4-3.Luc.R-E- (pNL4-3/Luc) (3 µg) and SARS-CoV-2 S expression plasmid (1 µg). Two days post-transfection, cell and virus lysates were prepared and subjected to Western blot analysis with anti-HIV-1 Ig to detect Gag proteins, anti-HIV-1 gp120, anti-HIV-1 gp41, anti-MLV gp70, anti-MLV p15(E), anti-SARS-CoV/SARS-CoV-2 S, anti-GBP5, or anti-alpha-tubulin. Positions of the HIV-1 Env glycoprotein precursor gp160, HIV-1 surface glycoprotein gp120, HIV-1 transmembrane glycoprotein gp41, Gag precursor Pr55Gag, p24 CA protein, MLV Env precursor Pr85Env, MLV surface Env glycoprotein gp70, MLV transmembrane protein p15(E), S precursor S0, and transmembrane spike glycoprotein S2 are indicated. The mobility of molecular mass standards is shown on the left of each blot in kilodaltons. The virion-associated levels of ( D ) HIV-1 gp120, ( E ) MLV gp70, and ( F ) SARS-CoV-2 S were quantified and normalized to virion-associated p24; values were set at 100% in the absence of GBP5. Analysis was performed with ImageJ 1.53k software. Data shown are means ± SDs from three to four independent experiments. Statistical significance (two-tailed unpaired t -test): * P < 0.05, ** P < 0.02, *** P < 0.01. ns, not significant.

Article Snippet: The following antibodies were used in this study: anti-HIV-1 Ig, anti-HIV-1 gp120 (16H3, catalog no. 12559), and anti-HIV-1 gp41 (clone Chessie 8, catalog no. ARP-13049) (obtained from the NIH ARP); anti-SARS-CoV/SARS-CoV-2 S (GeneTex, catalog no. GTX632604); anti-VSV-G (Cell Signaling, catalog no. 93372); anti-GBP5 (Cell Signaling, catalog no. 67798); anti-STAT-1 (Cell Signaling, catalog no. 14994); anti-alpha-tubulin (Sigma-Aldrich, clone B-5-1-2, catalog no. T5168); anti-CD4 (Santa Cruz, catalog no. sc-19641); goat anti-mouse IgG conjugated to horseradish peroxidase (HRP) (Invitrogen, catalog no. 31446); goat anti-rabbit IgG conjugated to HRP (Invitrogen, catalog no. 65-6120); and anti-human IgG conjugated to HRP (Sigma-Aldrich, catalog no. GENA933).

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Luciferase, Derivative Assay, Virus, Western Blot, Software, Two Tailed Test

Journal: mBio

Article Title: Guanylate-binding protein 5 antagonizes viral glycoproteins independently of furin processing

doi: 10.1128/mbio.02086-24

Figure Lengend Snippet: GBP5 reduces cell-surface HIV-1 Env and SARS-CoV-2 S expression. HEK293T cells were co-transfected with either wild-type (WT) GBP5 expression plasmid (0.5 µg) or the isoprenylation-deficient GBP5 mutant C583A (0.5 µg) and ( A ) the pNL4-3 molecular clone (4 µg) or ( B ) the Env(−) pNL4-3 derivative pNL4-3/KFS (3 µg) and SARS-CoV-2 S expression plasmid (1 µg). Two days post-transfection, cells were collected, incubated with anti-HIV-1 gp120 or Alexa Fluor 647-conjugated SARS-CoV-2 spike S1 subunit antibody, fixed, and permeabilized. After permeabilization, cells were incubated with goat anti-human antibody conjugated to Alexa Fluor 647 to stain HIV-1 gp120 and with PE-conjugated anti-HA.11 epitope tag antibody to detect GBP5. Histograms of HIV-1 Env- and SARS-CoV-2 S-positive Alexa Fluor 647 (AF647) expression were plotted in the absence of GBP5 (green) or in the presence of either C583A GBP5 (orange) or WT GBP5 (gray). Histogram of AF647 positivity of unstained cells (blue) was included as a control. Mean and median fluorescence intensity (MFI) of HIV-1 Env and SARS-CoV-2 S cell-surface expression was calculated with FlowJo version 10.9.0 software. The MFI in the absence of GBP5 is set at 100%. Data shown are means ± SDs from three independent experiments. Statistical significance (two-tailed unpaired t -test): *** P < 0.01; **** P < 0.0001. ns, not significant.

Article Snippet: The following antibodies were used in this study: anti-HIV-1 Ig, anti-HIV-1 gp120 (16H3, catalog no. 12559), and anti-HIV-1 gp41 (clone Chessie 8, catalog no. ARP-13049) (obtained from the NIH ARP); anti-SARS-CoV/SARS-CoV-2 S (GeneTex, catalog no. GTX632604); anti-VSV-G (Cell Signaling, catalog no. 93372); anti-GBP5 (Cell Signaling, catalog no. 67798); anti-STAT-1 (Cell Signaling, catalog no. 14994); anti-alpha-tubulin (Sigma-Aldrich, clone B-5-1-2, catalog no. T5168); anti-CD4 (Santa Cruz, catalog no. sc-19641); goat anti-mouse IgG conjugated to horseradish peroxidase (HRP) (Invitrogen, catalog no. 31446); goat anti-rabbit IgG conjugated to HRP (Invitrogen, catalog no. 65-6120); and anti-human IgG conjugated to HRP (Sigma-Aldrich, catalog no. GENA933).

Techniques: Expressing, Transfection, Plasmid Preparation, Mutagenesis, Incubation, Staining, Control, Fluorescence, Software, Two Tailed Test

Journal: mBio

Article Title: Guanylate-binding protein 5 antagonizes viral glycoproteins independently of furin processing

doi: 10.1128/mbio.02086-24

Figure Lengend Snippet: Overexpression of HIV-1 Env and SARS-CoV-2 S confers GBP5 resistance. HEK293T cells were co-transfected with either WT GBP5 expression plasmid (0.5 µg) or the isoprenylation-deficient GBP5 mutant C583A (0.5 µg) and the Env-defective pNL4-3 derivative pNL4-3/KFS and HIV-1 Env expression plasmid at a ratio of 1:10 ( A and B ) or 1:3 ( C and D ), or Env(−) luciferase-encoding NL4-3-derived vector pNL4-3.Luc.R-E- (pNL4-3/Luc) and SARS-CoV-2 S expression plasmid at a ratio of 1:5 ( E and F ) or 1:1 ( G and H ). Two days post-transfection, cells were collected, incubated with anti-HIV-1 gp120 or AF647-conjugated SARS-CoV-2 spike S1 subunit antibody, fixed, and permeabilized. After permeabilization, cells were incubated with goat anti-human antibody conjugated to AF647 to stain HIV-1 gp120 and with PE-conjugated anti-HA.11 epitope tag antibody to detect GBP5. Histograms of HIV-1 Env- ( A and C ) and SARS-CoV-2 S- ( E and G ) positive expression were plotted in the absence of GBP5 (green) or in the presence of either C583A GBP5 (orange) or WT GBP5 (gray). Histogram of AF647 positivity of unstained cells (blue) was included as a control. Mean and median fluorescence intensity (MFI) of HIV-1 Env and SARS-CoV-2 S cell-surface expression was calculated with FlowJo 10.9.0 software. The MFI in the absence of GBP5 is set at 100%. Data shown are means ± SDs from three independent experiments. Statistical significance (two-tailed unpaired t -test): * P < 0.05; ** P < 0.02; **** P < 0.0001. Virus supernatants were collected, normalized for RT, and used to infect TZM-bl cells ( B and D ) or HEK293T cells stably expressing hACE2 and transfected with TMPRSS2 expression plasmid ( F and H ). The infectivity in the absence of GBP5 is set at 100%. Data shown are means ± SDs from three independent experiments. Statistical significance (two-tailed unpaired t -test): *** P < 0.01; **** P < 0.0001. ns, not significant.

Article Snippet: The following antibodies were used in this study: anti-HIV-1 Ig, anti-HIV-1 gp120 (16H3, catalog no. 12559), and anti-HIV-1 gp41 (clone Chessie 8, catalog no. ARP-13049) (obtained from the NIH ARP); anti-SARS-CoV/SARS-CoV-2 S (GeneTex, catalog no. GTX632604); anti-VSV-G (Cell Signaling, catalog no. 93372); anti-GBP5 (Cell Signaling, catalog no. 67798); anti-STAT-1 (Cell Signaling, catalog no. 14994); anti-alpha-tubulin (Sigma-Aldrich, clone B-5-1-2, catalog no. T5168); anti-CD4 (Santa Cruz, catalog no. sc-19641); goat anti-mouse IgG conjugated to horseradish peroxidase (HRP) (Invitrogen, catalog no. 31446); goat anti-rabbit IgG conjugated to HRP (Invitrogen, catalog no. 65-6120); and anti-human IgG conjugated to HRP (Sigma-Aldrich, catalog no. GENA933).

Techniques: Over Expression, Transfection, Expressing, Plasmid Preparation, Mutagenesis, Luciferase, Derivative Assay, Incubation, Staining, Control, Fluorescence, Software, Two Tailed Test, Virus, Stable Transfection, Infection

Journal: mBio

Article Title: Guanylate-binding protein 5 antagonizes viral glycoproteins independently of furin processing

doi: 10.1128/mbio.02086-24

Figure Lengend Snippet: GBP5 reduces the glycosylation of HIV-1 Env, MLV Env, SARS-CoV-2 S, SARS-CoV S, and VSV-G. HEK293T cells were co-transfected with varying amounts of a GBP5 expression vector, with the total DNA held constant with empty vector (0.5 µg total), and ( A ) pNL4-3 (4 µg), ( B ) the Env(−) pNL4-3 derivative pNL4-3/KFS (3 µg) and MLV Env expression plasmid (0.6 µg), ( C ) Env(−) luciferase-encoding NL4-3-derived vector pNL4-3.Luc.R-E- (pNL4-3/Luc) (3.3 µg) and SARS-CoV-2 S expression plasmid (1 µg), ( D ) Env(−) luciferase-encoding NL4-3-derived vector pNL4-3.Luc.R-E- (pNL4-3/Luc) (3.3 µg) and SARS-CoV S expression plasmid (1 µg), or ( E ) the Env(−) pNL4-3 derivative pNL4-3/KFS (4 µg) and VSV-G expression plasmid (0.5 µg). Two days post-transfection, cell lysates were prepared and treated with PNGase F before being subjected to Western blot analysis with anti-HIV-1 gp120, anti-HIV-1 gp41, anti-MLV gp70, anti-SARS-CoV/SARS-CoV-2 S, anti-VSV-G, anti-GBP5, or anti-alpha-tubulin. The positions of the glycan-stripped viral glycoproteins in the PNGaseF-treated samples are indicated. The mobility of molecular mass standards is shown on the left of each blot in kilodalton.

Article Snippet: The following antibodies were used in this study: anti-HIV-1 Ig, anti-HIV-1 gp120 (16H3, catalog no. 12559), and anti-HIV-1 gp41 (clone Chessie 8, catalog no. ARP-13049) (obtained from the NIH ARP); anti-SARS-CoV/SARS-CoV-2 S (GeneTex, catalog no. GTX632604); anti-VSV-G (Cell Signaling, catalog no. 93372); anti-GBP5 (Cell Signaling, catalog no. 67798); anti-STAT-1 (Cell Signaling, catalog no. 14994); anti-alpha-tubulin (Sigma-Aldrich, clone B-5-1-2, catalog no. T5168); anti-CD4 (Santa Cruz, catalog no. sc-19641); goat anti-mouse IgG conjugated to horseradish peroxidase (HRP) (Invitrogen, catalog no. 31446); goat anti-rabbit IgG conjugated to HRP (Invitrogen, catalog no. 65-6120); and anti-human IgG conjugated to HRP (Sigma-Aldrich, catalog no. GENA933).

Techniques: Glycoproteomics, Transfection, Expressing, Plasmid Preparation, Luciferase, Derivative Assay, Western Blot

Journal: mBio

Article Title: Guanylate-binding protein 5 antagonizes viral glycoproteins independently of furin processing

doi: 10.1128/mbio.02086-24

Figure Lengend Snippet: GBP5 does not impair the processing of HIV-1 Env, MLV Env, or SARS-CoV-2 S glycoproteins under the conditions tested. Quantification of the ratios of cell-associated (A) HIV-1 gp160/gp120, (B) MLV Pr85Env/gp70, and (C) SARS-CoV-2 S0/S2 in the presence of varying amounts of GBP5 expression plasmid (0–0.5 µg) is shown. Quantification included only fully glycosylated forms of gp120, gp70, or S2 (dark bars) or fully glycosylated plus the more rapidly migrating, less-glycosylated forms (light bars). Blots below the graphs are reproduced from for ease of viewing; fully glycosylated forms of gp120, gp70, and S2 are denoted by #; lower-molecular-weight (less glycosylated) species are indicated by ##. Quantification of the ratios of cell-associated ( D ) HIV-1 gp160/gp120, ( E ) MLV Pr85Env/gp70, and ( F ) SARS-CoV-2 S0/S2 from PNGaseF-treated samples in the absence or presence of GBP5 expression plasmid (0.5 µg). Blots are reproduced from for ease of viewing, with the bands quantified in the graphs boxed in the blots. Analysis was performed with ImageJ 1.53k software. Glycoprotein processing in the absence of GBP5 is set at 100%. Data shown are means ± SDs from three to four independent experiments. Statistical significance (two-tailed unpaired t -test): * P < 0.05; ** P < 0.02; *** P < 0.01; **** P < 0.0001. ns, not significant.

Article Snippet: The following antibodies were used in this study: anti-HIV-1 Ig, anti-HIV-1 gp120 (16H3, catalog no. 12559), and anti-HIV-1 gp41 (clone Chessie 8, catalog no. ARP-13049) (obtained from the NIH ARP); anti-SARS-CoV/SARS-CoV-2 S (GeneTex, catalog no. GTX632604); anti-VSV-G (Cell Signaling, catalog no. 93372); anti-GBP5 (Cell Signaling, catalog no. 67798); anti-STAT-1 (Cell Signaling, catalog no. 14994); anti-alpha-tubulin (Sigma-Aldrich, clone B-5-1-2, catalog no. T5168); anti-CD4 (Santa Cruz, catalog no. sc-19641); goat anti-mouse IgG conjugated to horseradish peroxidase (HRP) (Invitrogen, catalog no. 31446); goat anti-rabbit IgG conjugated to HRP (Invitrogen, catalog no. 65-6120); and anti-human IgG conjugated to HRP (Sigma-Aldrich, catalog no. GENA933).

Techniques: Expressing, Plasmid Preparation, Molecular Weight, Software, Two Tailed Test

Journal: bioRxiv

Article Title: A hollow fiber membrane-based liver organoid-on-a-chip model for examining drug metabolism and transport

doi: 10.1101/2024.08.12.607504

Figure Lengend Snippet: (A) A representative whole-mount confocal immunofluorescence microscopy image of OATP1B3, OCT1, MDR1, MRP2 and MRP3 in a dorsal section of a HFM. (B) A representative confocal immunofluorescence microscopy image for the basolateral localization of NTCP in cell membranes of a monolayer in an anterior section of a HFM. (C) Representative brightfield images and images of intracellular fluorescence of calcein of monolayers treated with efflux inhibitors versus DMSO (control). (D) The quantification of calcein fluorescence. Data information: scale bars in A-C, 100 µm. In A and B, nuclei were counterstained with DAPI. In D, the graph shows mean ± SD (error bars). In A and B, three independent replicates (donors; 2, 3, 5) with three technical replicates (monolayers) were examined. In C and D, one independent replicate (donor, 5) with 40 technical replicates (fields) from two monolayers (20 replicates each) was examined.

Article Snippet: These included rabbit anti-ZO1 (1:50, Thermo Fisher Scientific, 40-2300), rabbit anti-OATP1B3 (1:50, Novus Biologicals, NBP1-80980), rabbit anti-OCT1 (1:50, Novus Biologicals, NBP1-59464), mouse anti-MDR1 (1:50, Santa Cruz Biotechnology, sc-55510), rabbit anti-MRP2 (1:50, Abcam, ab187644), rabbit anti-MRP3 (1:50, Novus Biologicals, NBP2-37923) and phalloidin (a toxin against F-actin) conjugated to Alexa Fluor 488 (1:400, Thermo Fisher Scientific, A12379).

Techniques: Immunofluorescence, Microscopy, Fluorescence, Control