Journal: Gene

Article Title: Period circadian regulator 2 suppresses drug resistance to cisplatin by PI3K/AKT pathway and improves chronochemotherapeutic efficacy in cervical cancer.

doi: 10.1016/j.gene.2021.146003

Figure Lengend Snippet: Fig. 5. Overexpression of PER2 regulates clock circadian and ameliorates drug resistance through PI3K/AKT pathway in DDP-resistant cervical cancer cells. (A) Representative images of western blot. (B-M) Quantitative results of (B, C) PER2, (D, E) CLOCK, (F, G) BMAL1, (H, I) CRY1, (J, K) MRP1 and (L, M) MDR1 in Hela/ DDP as well as SiHa/DDP cells with pcDNA3.1-PER2 transfection and/or hEGF treatment. **P < 0.01; ****p < 0.0001.

Article Snippet: The sample was transferred onto the membrane at a gel volume of 1.5 mA/cm2 for 1.5 h. The membrane was added with 5% skimmed milk powder + TBST, and shaken in a shaker at room temperature lasting 1 h. The membranes were added with primary antibodies against PER2 (67513-1-Ig; Proteintech), CLOCK (18094-1-AP; Proteintech), BMAL1 (14268-1-AP; Proteintech), CRY1 (13474-1-AP; Proteintech), MRP1 (67228–1-Ig; Proteintech), MDR1 (22336-1-AP; Proteintech), PI3K (20584-1-AP; Proteintech), p-PI3K (17366S; CST), AKT (10176-2-AP; Proteintech), p-AKT (66444-1-Ig; Proteintech), Snail1 (13099-1-AP; Proteintech), Twist (25465–1-AP; Proteintech), E-cadherin (20874-1- AP; Proteintech), Vimentin (10366-1-AP; Proteintech) and GAPDH (60004-1-Ig; Proteintech) which was diluted at 1:1000 with 5% BSA + TBS, and shaken overnight at 4 ◦C.

Techniques: Over Expression, Western Blot, Transfection

Journal: Gene

Article Title: Period circadian regulator 2 suppresses drug resistance to cisplatin by PI3K/AKT pathway and improves chronochemotherapeutic efficacy in cervical cancer.

doi: 10.1016/j.gene.2021.146003

Figure Lengend Snippet: Fig. 9. Cisplatin therapy at the peak of PER2 expression ameliorates chemotherapy resistance and EMT in cervical cancer. (A) Representative images of western blot. Assessment of the expression of (B) PER2, (C) CLOCK, (D) BMAL1, (E) CRY1, (F) MRY1, (G) MRP1, (H) PI3K, (I) p-PI3K, (J) p-PI3K/PI3K, (K) AKT, (L) p-AKT, (M) p- AKT/AKT, (N) Snail, (O) Twist, (P) Vimentin and (Q) E-cadherin in tumor tissues from Hela/DDP cells-induced nude mice treated with Cisplatin at the peak or trough of PER2 expression. *P < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The sample was transferred onto the membrane at a gel volume of 1.5 mA/cm2 for 1.5 h. The membrane was added with 5% skimmed milk powder + TBST, and shaken in a shaker at room temperature lasting 1 h. The membranes were added with primary antibodies against PER2 (67513-1-Ig; Proteintech), CLOCK (18094-1-AP; Proteintech), BMAL1 (14268-1-AP; Proteintech), CRY1 (13474-1-AP; Proteintech), MRP1 (67228–1-Ig; Proteintech), MDR1 (22336-1-AP; Proteintech), PI3K (20584-1-AP; Proteintech), p-PI3K (17366S; CST), AKT (10176-2-AP; Proteintech), p-AKT (66444-1-Ig; Proteintech), Snail1 (13099-1-AP; Proteintech), Twist (25465–1-AP; Proteintech), E-cadherin (20874-1- AP; Proteintech), Vimentin (10366-1-AP; Proteintech) and GAPDH (60004-1-Ig; Proteintech) which was diluted at 1:1000 with 5% BSA + TBS, and shaken overnight at 4 ◦C.

Techniques: Expressing, Western Blot

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Identification of microRNAs and mRNAs associated with multidrug resistance of human laryngeal cancer Hep-2 cells

doi: 10.1590/1414-431X20131662

Figure Lengend Snippet: Rhodamine 123 efflux is increased and multidrug resistance 1 (MDR1) expression is up-regulated in Hep-2/v cells compared to Hep-2 cells. A , Rhodamine 123 retention in Hep-2 and Hep-2/v cells. The cells were incubated with rhodamine 123 and measured by flow cytometry. Data are reported as mean±SD percentages of rhodamine 123-positive cells. *P<0.01 vs Hep-2 cells (chi-square test). B , Expression of MDR1 mRNA in Hep-2 and Hep-2/v cells. The relative levels of MDR1 mRNA in Hep-2 and Hep-2/v cells were determined by real-time reverse transcription-polymerase chain reaction. Data are reported as means±SD. *P<0.01 vs Hep-2 cells (Student t -test). C , Expression of the MDR1 protein in Hep-2 and Hep-2/v cells. The relative levels of the MDR1 protein in Hep-2 and Hep-2/v cells were determined by Western blotting. Data are reported as means±SD. *P<0.01 vs Hep-2 cells (Student t -test).

Article Snippet: The membrane was blocked for 1'h at room temperature in PBS containing 0.3% Tween 20 and 5% skim milk and incubated overnight at 4ºC with an anti-MDR1 antibody (dilution 1:1,000; Chemicon, USA) or anti-actin antibody (dilution 1:1,500; Chemicon).

Techniques: Expressing, Incubation, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: List of antibodies used in the experiments.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques:

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: Primers used for detection of multidrug resistance proteins.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: Antibody concentrations used for Western blotting analysis of multidrug resistance protein expressed by endothelial cells.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: Antibody concentrations used for flow cytometry analysis of multidrug resistance proteins expressed by endothelial cells.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Flow Cytometry

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: Substrates and inhibitors used in functional tests.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Functional Assay

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: Flow cytometry analysis of multidrug resistance protein expressed by endothelial cells.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Flow Cytometry

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: MDR1, MRP1 and BCRP protein levels were revealed with different sets of specific antibodies in endothelial cell extracts obtained lysing cells with RIPA buffer. Protein extracts from LoVo/Dx cells were analyzed as positive controls. MDR1/4: 180 kDa; MDR1/5: 170 kDa, MRP1/3: 180 kDa; MRP1/4: 170–220 kDa; MRP1/5: 190 kDa; BCRP/2: 72 kDa; BCRP/3 65–80 kDa and BCRP/4: 72 kDa. Equal protein loading (50 μg/line) was confirmed by β-actin expression (45 kDa).

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: A challenging correlation

doi: 10.1371/journal.pone.0172371

Figure Lengend Snippet: Summary of results. Numbers indicate positive (1) or negative (0) results obtained by particular technique used. In case of ICC the gradation of results is presented as (+) and (-) score. M- mRNA; WB- Western Blotting FC- Flow Cytometry; ICC- immunocytochemistry; FA- standard functional test; FB- commercial functional test. FA* and FB* refers to functional tests applied for the whole MRP protein family. All tests were repeated at least three times, and both functional tests were each time performed in triplicate.

Article Snippet: Anti-MDR1/ABCB1 , MDR1/1 , 15D3 , external , PE , FC , BD Pharmingen.

Techniques: Western Blot, Flow Cytometry, Immunocytochemistry, Functional Assay

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