Journal: Journal of Veterinary Pharmacology and Therapeutics

Article Title: Comparison of Human and Canine P‐Glycoprotein Substrates at R, P, and H Binding Sites

doi: 10.1111/jvp.70034

Figure Lengend Snippet: Western blots assessing MRP1 (~190 kDa; top left panel) and P‐gp (~170 kDa; top right panel) expression in various cell lines. Beta actin (~42 kDa; middle panels) was used as a loading control. Molecular weight markers (MW: 250, 150, 50, 37 kDa) are indicated along the left side of the respective images. Normalized MRP1 and P‐gp expression are indicated in the bottom panels. HeLa cells are the positive control cell line for MRP1 expression (see Note 2); K9KO are MDCK cells with canine P‐gp knocked out (Karlgren et al. ), MDCK cells, Madin Darby canine kidney cell line; hMDR1 over‐expresses human P‐gp (Karlgren et al. ); K9PGP cells overexpress canine P‐gp (Mealey et al. ).

Article Snippet: The fluorescent multidrug resistance protein 1 (MRP1) substrate 5(6)‐carboxyfluorescein diacetate (CFDA) was purchased from Thermo Fisher (Eugene, OR) and the MRP1 specific inhibitor MK 571 was obtained from Tocris Bioscience (Minneapolis, MN).

Techniques: Western Blot, Expressing, Control, Molecular Weight, Positive Control

Journal: Biomedical Reports

Article Title: Scutellarein enhances cisplatin‑induced apoptotic effects by suppressing the PI3K/AKT‑MDR1 pathway in human NPC/HK1 nasopharyngeal carcinoma cells

doi: 10.3892/br.2025.1938

Figure Lengend Snippet: Scutellarein enhances cisplatin-induced inhibition of cell viability and release of cytokeratin 18 fragments by inhibiting the PI3K/AKT-MDR1 pathway in NPC/HK1 cells. (A) NPC/HK1 cells were treated without (Ctrl) or with 12.5 µM scutellarein, 4.15 µM cisplatin, or their combination for 48 h. Protein expression of p-AKT, AKT, MDR1, and GAPDH was examined using an immunoblotting assay. (B) NPC/HK1 cells were treated without (Ctrl) or with 4.15 µM cisplatin, or 4.15 µM cisplatin plus 10 µM LY294002 for 48 h. Upper panel, protein expression of p-AKT, AKT, MDR1, and GAPDH was examined using an immunoblotting assay. Lower panel, cytokeratin 18 fragment levels in the cell culture supernatant were measured by ELISA. (C) NPC/HK1 cells were treated without (Ctrl) or with 4.15 µM cisplatin, or 4.15 µM cisplatin plus 10 µM PSC833 for 48 h. Upper panel, protein expression of MDR1 was examined using an immunoblotting assay. Middle panel, cell viability was assessed using the MTT assay. Lower panel, cytokeratin 18 fragment levels in the cell culture supernatant were measured by ELISA. Statistical significance was indicated as follows: * P<0.05 vs. Ctrl; and # P<0.05 vs. Cis alone; n=3. p-, phosphorylated; MDR1, multidrug resistance protein 1; Ctrl, control; Scu, scutellarein; Cis, cisplatin; LY, LY294002.

Article Snippet: Primary antibodies targeting caspase-8 (cat. no. 9746; 1:1,000), cleaved caspase-8 (cat. no. 9429; 1:1,000), caspase-9 (cat. no. 9502), cleaved caspase-9 (cat. no. 9505), caspase-7 (cat. no. 9492; 1:1,000), cleaved caspase-7 (cat. no. 9491; 1:1,000), cleaved poly (ADP-ribose) polymerase (PARP) (cat. no. 9541; 1:1,000), phosphorylated (p)-AKT (cat. no. 9271; 1:1,000), AKT (cat. no. 9272; 1:1,000), Beclin 1 (cat. no. 3738; 1:1,000), multidrug resistance protein 1 (MDR1) (cat. no. 13342; 1:1,000), and GAPDH (cat. no. 97166; 1:5,000) were purchased from Cell Signaling Technology, Inc.

Techniques: Inhibition, Expressing, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, MTT Assay, Control

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New uracil analog as inhibitor/modulator of ABC transporters or/and NF-κB in taxol-resistant MCF-7/Tx cell line

doi: 10.1007/s00432-024-05833-z

Figure Lengend Snippet: Primer sequences for real-time PCR reaction

Article Snippet: The protein levels of ABCB1 and ABCG2 in MCF-7 and MCF-7/Tx cells were determined using an ELISA-based method, including ABCB1 (Multidrug resistance protein 1 kit) and ABCG2 (ATP-binding cassette sub-family G member 2 kit), provided by Finetest BT LAB (Shanghai, China).

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New uracil analog as inhibitor/modulator of ABC transporters or/and NF-κB in taxol-resistant MCF-7/Tx cell line

doi: 10.1007/s00432-024-05833-z

Figure Lengend Snippet: Real-time PCR analysis of mRNA levels of ABCB1, ABCG2, and NF-κB in MCF-7/Tx cells compared to parental MCF-7 cells ( A ). The effect of Tx, U-359, or a combination of U-359 and Tx treatment on mRNA expression levels of ABCB1 ( B ), ABCG2 ( C ), and NF-κB ( D ). Data are presented as mean ± SEM. In each experiment, three replicates were used. Statistical significance was evaluated using one-way ANOVA followed by a post-hoc multiple comparison Student–Newman–Keuls test. * p <0.05, *** p < 0.001, compared with control; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with Tx

Article Snippet: The protein levels of ABCB1 and ABCG2 in MCF-7 and MCF-7/Tx cells were determined using an ELISA-based method, including ABCB1 (Multidrug resistance protein 1 kit) and ABCG2 (ATP-binding cassette sub-family G member 2 kit), provided by Finetest BT LAB (Shanghai, China).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Comparison, Control

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: New uracil analog as inhibitor/modulator of ABC transporters or/and NF-κB in taxol-resistant MCF-7/Tx cell line

doi: 10.1007/s00432-024-05833-z

Figure Lengend Snippet: The analysis of ABCB1 and ABCG2 protein changes in MCF-7/Tx cells compared to control (parental MCF-7 cells set at 100%) ( A ). The effect of Tx, U-359, and U-359+Tx treatments (each at IC 50 concentration) on the protein levels of ABCB1 in MCF-7 ( B ) and MCF-7/Tx ( C ) cells, and ABCG2 in MCF-7 ( D ) and MCF-7/Tx ( E ) cells. Data are presented as mean±SEM. In each experiment, three replicates were used. The control consisted of untreated MCF-7 cells. Statistical significance was assessed by one-way ANOVA and a post-hoc multiple comparison Student–Newman–Keuls test *** p < 0.01 in comparison with control; ### p < 0.01 in comparison with Tx

Article Snippet: The protein levels of ABCB1 and ABCG2 in MCF-7 and MCF-7/Tx cells were determined using an ELISA-based method, including ABCB1 (Multidrug resistance protein 1 kit) and ABCG2 (ATP-binding cassette sub-family G member 2 kit), provided by Finetest BT LAB (Shanghai, China).

Techniques: Control, Concentration Assay, Comparison