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: Human Gene Therapy

Article Title: Life-Long Correction of Hyperbilirubinemia with a Neonatal Liver-Specific AAV-Mediated Gene Transfer in a Lethal Mouse Model of Crigler–Najjar Syndrome

doi: 10.1089/hum.2013.233

Figure Lengend Snippet: Tissue distribution of bilirubin transporters. (A) RT-PCR of total liver from WT mice, AAT-hUGT1A1-treated mutant mice (MUT AAT), and skeletal muscle of CMV-hUGT1A1-treated mutant mice (MUT CMV). Mouse Gapdh was used as endogenous control. (B) Western blot analysis using anti-Mrp2 and anti-Mrp3 antibodies of total liver and skeletal muscle protein extracts (50 μg) from WT, mutant treated with AAT-hUGT1A1 (liver), or CMV-hUGT1A1 (SM, skeletal muscle). β-tubulin was used as loading control.

Article Snippet: Primary antibodies used were as follows: anti-human UGT1 rabbit polyclonal antibody, anti-Mrp2 and anti-Mrp3 (Santa Cruz Biotechnology), and anti-β-tubulin mAb E7 (Developmental Studies Hybridoma Bank).

Techniques: Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Western Blot

Journal: Biomicrofluidics

Article Title: Bile canaliculi formation by aligning rat primary hepatocytes in a microfluidic device

doi: 10.1063/1.3580753

Figure Lengend Snippet: Results of the immunostaining. (a) The distribution of apical marker MRP2 protein (green fluorescence). MRP2 was concentrated in the localized regions along the cell-cell contacts. (b) The distribution of basolateral marker CD147 protein (red fluorescence). CD147 was expressed in almost all the areas across the channel corresponding to the locations of cell membranes.

Article Snippet: The cells were fixed with 4% paraformaldehyde in PBS for 15 min and then were blocked with 0.05% Triton X-100 and 1% bovine serum albumin in PBS at room temperature for 1 h. Next, the cells were incubated with a primary mouse anti-MRP2 monoclonal antibody (Enzo), a mouse anti-CD147 monoclonal antibody (AbD Serotec), and a rabbit ZO-1 polyclonal antibody (Invitrogen) for 12 h at room temperature.

Techniques: Immunostaining, Marker, Fluorescence

Journal: Oncology Letters

Article Title: Overexpression of close homolog of L1 enhances the chemosensitivity of lung cancer cells via inhibition of the Akt pathway

doi: 10.3892/ol.2020.11972

Figure Lengend Snippet: CHL1 is downregulated in DDP and PTX-resistant A549 cells. (A) Cell survival of A549 and A549-resistant cells (A549/DDP and A549/PTX) treated with increasing concentrations of DDP and PTX, as assessed by MTT assay. (B) The IC 50 values of DDP in A549/DDP and A549 cells, and the IC 50 values of PTX in A549/PTX and A549 cells. *P<0.05 vs. A549 cells. (C) Western blotting demonstrated the expression of drug resistance-related proteins MDR1, MRP and LRP in A549 cells and A549-resistant cells (A549/DDP and A549/PTX). *P<0.05 vs. A549 cells. The protein and mRNA expression levels of CHL1 in A549 cells and A549-resistant cells (A549/DDP and A549/PTX) were analysed by (D) western blotting and (E) reverse transcription-quantitative PCR, respectively. *P<0.05 vs. A549 cells. (F) The mRNA expression of CHL1 in H460 and H460/DDP cells in the GSE21656 dataset. *P<0.05 vs. H460 cells. CHL1, close homolog of L1; DDP, cisplatin; PTX, paclitaxel; MDR1, multi-drug resistance gene 1; MRP, multidrug resistance-associated protein; LRP, low-density lipoprotein receptor-related protein; IC50, half maximal inhibitory concentration.

Article Snippet: Next, the proteins were transferred onto a polyvinylidene membrane (Thermo Fisher Scientific, Inc.), blocked with 5% BSA (Thermo Fisher Scientific, Inc.) for 2 h at 4°C, and incubated overnight at 4°C with primary antibodies against CHL1 (1:500; cat. no. 25250-1-AP; ProteinTech, Inc.), multi-drug resistance gene 1 (MDR1; 1:500; cat. no. 22336-1-AP; ProteinTech, Inc.), multidrug resistance-associated protein (MRP; 1:500; cat. no. 67228-1-Ig; ProteinTech, Inc.), low-density lipoprotein receptor-related protein (LRP; 1:500; cat. no. 22336-1-AP; ProteinTech, Inc.), phosphorylated (p)-Akt (1:1,000; cat. no. ab38449; Abcam,) and Akt (1:2,000; cat. no. ab227385; Abcam).

Techniques: MTT Assay, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Concentration Assay