Journal: International Journal of Molecular Sciences

Article Title: Augmented Therapeutic Potential of EC-Synthetic Retinoids in Caco-2 Cancer Cells Using an In Vitro Approach

doi: 10.3390/ijms23169442

Figure Lengend Snippet: Western blotting analysis of ABCB1 ( MDR1 ), ABCC1 ( MRP1 ) and Hsp70 in Caco-2 cells. ( A ) Representative immunoblotting images demonstrate the effect of IC 50 dose of ATRA, EC19, and EC23 on the expression levels of the indicated proteins in Caco-2 cells treated for 24 h. The expression of the investigated proteins in treated Caco-2 cells was calculated and normalized to the β-actin protein level in each treated sample, and finally plotted as ( B ) the mean protein fold change from negative control (0.1%DMSO) (dotted line) ± SEM. It can be clearly seen that EC19 and E23 caused significant downregulation of all proteins in comparison to the 0.1% DMSO control. * p < 0.05, ** p < 0.01 and *** p < 0.001 indicate a statistically significant difference. ns; non-significant.

Article Snippet: The membranes were then blocked, washed and incubated with antibodies against MDR1 (1:1000, #12683; Cell Signaling Technology, Danvers, MA, USA), MRP1 (1: 1500, #72202, Cell Signaling Technology), Hsp70 (1:1000, #MA3-009, Thermo Scientific, Waltham, MA, USA) and β-actin (1:3000, #A5060, Sigma-Aldrich, St. Louis, MO, USA) for 16–17 h at 4 °C in a humidified chamber.

Techniques: Western Blot, Expressing, Negative Control, Comparison, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Babao Dan Reverses Multiple-Drug Resistance in Gastric Cancer Cells via Triggering Apoptosis and Autophagy and Inhibiting PI3K/AKT/mTOR Signaling

doi: 10.1155/2021/5631942

Figure Lengend Snippet: BBD increased doxorubicin and Rho123 accumulation and downregulated the protein expression of ABCB1, ABCC1, and ABCG2 in gastric cancer-resistant cells. SGC7901/DDP cells were treated with BBD (0.25, 0.5, or 1.0 mg/mL) for 48 h, doxorubicin and Rho123 staining were observed via fluorescence microscope ((a) and (c)), and the average fluorescence intensity of all the cells was calculated in five random fields ((b) and (d)). Expression and quantitative analyses of ABCB1, ABCC1, and ABCG2 protein are shown ((e) and (f)). ∗ P < 0.05 and ∗∗ P < 0.01.

Article Snippet: ABCB1 (cat no. 13978), ABCC1 (cat no. 72202), ABCG2 (cat. no. 42078), cleaved caspase-3 (cat. no. 9664), mammalian target of rapamycin (mTOR) (cat no. 2983), and phospho-mTOR (cat no. 5536) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Expressing, Staining, Fluorescence, Microscopy

Journal: Chemical research in toxicology

Article Title: ATF3 Modulates the Resistance of Breast Cancer Cells to Tamoxifen through an N 6 ‑Methyladenosine-Based Epitranscriptomic Mechanism

doi: 10.1021/acs.chemrestox.1c00206

Figure Lengend Snippet: ATF3 transcriptionally regulates ABCB1 in TamR MCF-7 cells. (A) Western blot images and (B) quantification results for examining the levels of ABCB1 protein in MCF-7 and TamR MCF-7 cells, TamR MCF-7 cells after knockdown of ATF3, or TamR MCF-7 cells upon the overexpression of YTHDF2 and its W432A mutant. The relative levels of ABCB1 protein were measured from band intensities using ImageJ and are depicted relative to the level detected for control cells. (C) Relative levels of ABCB1 mRNA in parental and TamR MCF-7 cells or in TamR MCF-7 cells after knockdown of ATF3. (D) TamR MCF-7 cells with a stable knockdown of ABCB1 were treated with different concentrations of 4-OHT for 72 h, and cell viability was measured using CCK8. The data were normalized against the control groups (ethanol) and represented as mean ± SD ( n = 3). (E) ChIP-qPCR data showed thw occupancy of ATF3 in the enhancer regions of ABCB1 genes in TamR MCF-7 cells. The p -values were calculated on the basis of unpaired, two-tailed Student’s t -test: *, 0.01 ≤ p < 0.05; **, 0.001 ≤ p < 0.01; ***, p < 0.001; ns, not significant (i.e., p > 0.05).

Article Snippet: Primary antibodies employed for Western blot analysis included those recognizing human ATF3 (Santa Cruz Biotechnology, sc-81189), YTHDF2 (EMD Millipore, Q2490423), ABCB1 (ABclonal, A11747), and AK4 (Santa Cruz Biotechnology, sc-271161).

Techniques: Western Blot, Knockdown, Over Expression, Mutagenesis, Control, ChIP-qPCR, Two Tailed Test

Journal: Viruses

Article Title: Hepatitis C Virus-Induced FUT8 Causes 5-FU Drug Resistance in Human Hepatoma Huh7.5.1 Cells

doi: 10.3390/v11040378

Figure Lengend Snippet: Inhibition of P-gp and MRP1 increased the 5-FU drug sensitivity after HCV infection. ( A , C ) Western blot analysis of P-gp and MRP1 expression in Huh7.5.1 cells in the presence or absence of NSC23925 (P-gp inhibitor) and MK571 (MRP1 inhibitor). Western blot quantification analysis is also listed as ( B , D ). ( E – H ) Analysis of the cytotoxicity of drugs (5-FU/MTX/ADR/CDDP) in HCVcc-infected Huh7.5.1 cells in the presence of P-gp and MRP1 inhibitors.

Article Snippet: P-glycoprotein (P-gp) inhibitor NSC23925 and multidrug resistance-associated protein-1 (MRP1) inhibitor MK571 were purchased from MedChemExpress (MCE) (Monmouth Junction, NJ, USA) and dissolved in dimethyl sulfoxide (DMSO).

Techniques: Inhibition, Infection, Western Blot, Expressing

Journal: Oncology Reports

Article Title: Antitumor activity of the dual PI3K/mTOR inhibitor gedatolisib and the involvement of ABCB1 in gedatolisib resistance in canine tumor cells

doi: 10.3892/or.2022.8272

Figure Lengend Snippet: ABCB1 inhibition enhances the antitumor activity of gedatolisib in LCTCC cells. (A) Representative histograms of rhodamine-123 uptake (red) and efflux (black) in the low (MegTCC) and high (LCTCC) IC 50 cell lines. The gray histograms indicate cell auto-fluorescence. (B) Percent decrease in rhodamine-123 intensity in canine tumor cell lines. Each plot represents the mean percent decrease of rhodamine-123 intensity from three independent experiments, with bars representing the median (five high IC50 and seven low IC 50 cell lines). (C) Histograms of rhodamine-123 uptake/efflux in the absence (control) or presence of 5 µM cyclosporin A treatment in LCTCC cells. (D) Dose-response curves of LCTCC cells treated with increasing concentrations of gedatolisib in the absence or presence of 5 µM cyclosporin A. The results are presented as the mean ± SD of five replicates. (E) Immunoblotting analysis of LCTCC cells treated with gedatolisib (100 nM), with or without cyclosporin A (5 µM). (F) Cell cycle analysis of LCTCC cells treated with gedatolisib (100 nM), cyclosporin A (5 µM), or both agents for 24 h. Data are presented as the mean ± SD from three independent experiments. **P<0.01 and ***P<0.001 (for B, Mann-Whitney U test; for F, multiple t-tests). ABCB1, ATP-binding cassette, subfamily B, member 1; p-, phosphorylated; p70S6K, p70S6 kinase.

Article Snippet: ABCB1 inhibitors, cyclosporin A and tariquidar, were purchased from Wako Pure Chemical Industries, Ltd. and MedChemExpress, respectively.

Techniques: Inhibition, Activity Assay, Fluorescence, Control, Western Blot, Cell Cycle Assay, MANN-WHITNEY, Binding Assay