Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

Article Title: Noninvasive and quantitative assessment of the function of multidrug resistance-associated protein 1 in the living brain.

doi: 10.1038/jcbfm.2008.135

Figure Lengend Snippet: Figure 2 Analysis of chemical form. HPLC analysis of brain tissue extract at 15 (diamond) and 60 (square) mins after the intravenous injection of [14C]7m6BP into Mrp1 KO (solid) and WT (open) mice. Data are shown from a single mouse for each time point and each group.

Article Snippet: Male FVB mice and Mrp1 KO mice were purchased from Clea Japan Inc. (Tokyo, Japan) and Taconic Farms Inc. (Germantown, NY, USA), respectively.

Techniques: Injection

Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

Article Title: Noninvasive and quantitative assessment of the function of multidrug resistance-associated protein 1 in the living brain.

doi: 10.1038/jcbfm.2008.135

Figure Lengend Snippet: Figure 3 Time–radioactivity curves in the brain obtained by animal PET after intravenous injection of [11C]7m6BP into Mrp1 KO (solid symbols) and WT mice (open symbols) (A). Values are given as the mean±s.d. The two curves are representatives from six independent experiments on each group of mice. Efflux rates of [11C]PSG estimated from the time–radioactivity curves (B). Values are given as the mean±s.d. (n = 6, each group). The inset represents the common logarithm of the brain radioactivity during the period of 15 to 60 mins after injection. There was significant difference in efflux rates between Mrp1 KO and WT mice (Wilcoxon’s rank sum test, W = 21, a = 0.01). ID: injected dose.

Article Snippet: Male FVB mice and Mrp1 KO mice were purchased from Clea Japan Inc. (Tokyo, Japan) and Taconic Farms Inc. (Germantown, NY, USA), respectively.

Techniques: Radioactivity, Injection

Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

Article Title: Noninvasive and quantitative assessment of the function of multidrug resistance-associated protein 1 in the living brain.

doi: 10.1038/jcbfm.2008.135

Figure Lengend Snippet: Figure 5 Putative pathway and mechanism for the efflux transport of PSG. PSG (circle-SG) will be generated within glial cells, neuron, or BCEC. Based on our previous and present studies, the efflux pathway of PSG via simple diffusion is negligible, and the elimination by bulk flow is relatively small. PSG formed within parenchyma cells is first transported to the extracellular space by MRP1, and then taken up into BCEC by as yet unknown transporter. Finally, PSG is extruded into the blood by MRP1 at the luminal side of BCEC. This diagram does not represent other transporters although these are possibly involved in PSG efflux out of the brain.

Article Snippet: Male FVB mice and Mrp1 KO mice were purchased from Clea Japan Inc. (Tokyo, Japan) and Taconic Farms Inc. (Germantown, NY, USA), respectively.

Techniques: Generated, Diffusion-based Assay

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: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: LTH-sEV promotes LSEC capillarization in HFD mice and aggravates the progression of MASLD. ( A ) NTA analysis of LTH-sEV; ( B ) TEM images of LTH-sEV, scale bar = 100 nm; ( C ) Western blot analysis of CD9, CD63, Alix, TSG101, and calnexin protein expression in hepatocyte (HepG2) and LTH-sEV; ( D ) Schematic diagram of LTH-sEV injection into mice on HFD and NCD diet; ( E ) In vivo imaging detection of DiR-labeled LTH-sEV in mice; ( F ) SEM images and porosity of hepatic sinusoidal in NCD-fed mice treated with PBS, LTH-sEV, HFD-fed mice treated with PBS, LTH-sEV, n = 6, scale bar = 500 nm; ( G ) Immunohistochemistry images of CD31, Ang-2 in each group, scale bar = 50 μm, n = 6; ( H ) ELISA analysis of serum Ang-2 in each group, n = 6; ( I ) Western blot analysis of Ang-2 expression in each group, n = 3; ( J ) H&E staining, immunohistochemistry images of α-SMA, Sirius red staining in each group, scale bar = 50 μm, n = 6; ( K ) Serum ALT, AST expression in each group, n = 6; ( L ) qRT-PCR analysis of IL-1β, IL-6, and TNFα expression in each group, n = 6; ( M ) ELISA analysis of IL-1β, IL-6, and TNFα level in mice liver or serum, n = 6. Compared with HFD or NCD group, ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ns, no significance.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Western Blot, Expressing, Injection, In Vivo Imaging, Labeling, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Staining, Quantitative RT-PCR

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: OGT level in serum and serum sEV is positively associated with MASLD. ( A ) ELISA analysis of serum OGT expression in healthy individuals and patients with MASLD, n = 50; ( B ) ROC curve of serum OGT, AUC = .6832, P = .0016, Compared with Healthy group, ∗∗ P < .01; ( C ) Correlation analysis of serum OGT and ALT, r = .2154, P = .0438; ( D ) Correlation analysis of serum OGT and AST, r = .2493, P = .0171; ( E ) Correlation analysis of serum OGT and TG, r = .2541, P = .0197; ( F ) Immunohistochemical images of OGT expression in liver of healthy individuals and patients with MASLD; ( G ) Schematic diagram of the collection of Healthy-sEV and MASLD-sEV; ( H–I ) NTA and TEM detection of Healthy-sEV and MASLD-sEV, scale bar = 100 nm; ( J ) Western blot analysis of CD9, CD63, Alix and TSG101 protein expression in Healthy-sEV and MASLD-sEV; ( K ) Western blot analysis of OGT protein expression in Healthy-sEV and MASLD-sEV, n = 3; ( L ) ELISA analysis of of OGT in Healthy-sEV and MASLD-sEV, n = 50, Compared with Healthy-sEV group, ∗∗ P < .01; ( M ) ROC curve of OGT in serum sEV, AUC = .7228, P = .0001; ( N ) Correlation analysis of sEV-OGT and serum ALT, r = .2395, P = .0246; ( O ) Correlation analysis of sEV-OGT and serum AST, r = .2493, P = .0171; ( P ) Correlation analysis of sEV-OGT and Fib-4 score (Fib-4 = age × AST/PLT × √ALT), r = .3100, P = .0113.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemical staining, Western Blot

Journal: Biology of reproduction

Article Title: CD9 is a surface marker on mouse and rat male germline stem cells.

doi: 10.1095/biolreprod.103.020867

Figure Lengend Snippet: FIG. 1. Expression of CD9 in mouse testis. A) Characterization of anti-CD9 antibody-purified mouse testis cells using flow cytometry. Unselected (upper row) and anti-CD9 antibody-selected (lower row) testis cells were stained with FITC-conjugated streptavidin, PE-conjugated anti-a6-integrin antibody, or APC-conjugated anti-c-kit antibody, and the fluorescence was compared with that of controls. The dotted lines show the range of fluores- cence in the controls. Note the increased percentages of all three antigens in the anti-CD9 antibody-selected cell population. B) Immunohistological staining of CD9 antigen in mouse testis. Positive cells were found on the basement membrane of seminiferous tubules (arrows). Staining was also observed in the interstitial cells. Bar 5 50 mm (section). Stain: DAB followed by hematoxylin.

Article Snippet: Mouse anti-rat CD9 antibody was used with goat anti-mouse IgG microbeads (25 ml; Miltenyi Biotec) to select cells expressing rat CD9 molecules.

Techniques: Expressing, Cytometry, Staining, Membrane

Journal: Biology of reproduction

Article Title: CD9 is a surface marker on mouse and rat male germline stem cells.

doi: 10.1095/biolreprod.103.020867

Figure Lengend Snippet: FIG. 2. Expression of CD9 on mouse spermatogonial stem cells. A) Gross and histological appearance of recipient testes after transplantation of LacZ-marked donor testis cells. Left: Testis from a W recipient mouse 2 mo after transplantation of anti-CD9 antibody-selected cells. Middle: Testis from a W recipient mouse 2 mo after transplantation of unselected testis cells. Right: Histological section from a W recipient testis 2 mo after transplantation of anti-CD9-antibody-selected testis cells. Note the normal appearance and organization of spermatogenesis. Stain: X-gal followed by hematoxylin and eosin. B) Enhanced colonization of recipient testis by anti-CD9 antibody-selected mouse testis cells. The degree of coloni- zation in three experiments is indicated by the number of individual blue colonies. The values (mean 6 SEM) for anti-CD9 antibody-selected cells and unselected cells were 5.5 6 0.8 (n 5 16) and 0.8 6 0.2 (n 5 18) cells per 3 3 104 injected cells, respectively. Bars 5 1 mm (testes) and 50 mm (section).

Article Snippet: Mouse anti-rat CD9 antibody was used with goat anti-mouse IgG microbeads (25 ml; Miltenyi Biotec) to select cells expressing rat CD9 molecules.

Techniques: Expressing, Transplantation Assay, Staining, Injection

Journal: Biology of reproduction

Article Title: CD9 is a surface marker on mouse and rat male germline stem cells.

doi: 10.1095/biolreprod.103.020867

Figure Lengend Snippet: FIG. 3. Expression of CD9 on rat spermatogonial stem cells. A) Characterization of anti-CD9 antibody-purified rat testis cells using flow cytometry. Unselected (left) and anti-CD9 antibody-selected (right) testis cells were stained with Cy5-conjugated anti-mouse IgG antibody, and the fluorescence was compared with similar cell controls. The dotted lines show the control range of fluorescence. Note the increase in percentage of CD9-positive cells after magnetic selection. B) The appearance of recipient testes after transplantation of green fluorescent protein-marked donor testis cells. Seminiferous tubules were dissected using fine forceps. Left: Testis from a busulfan-treated nude recipient mouse 3 mo after transplantation of unselected testis cells (8.2 3 104 cells injected). Right: Testis from a busulfan-treated nude recipient mouse 3 mo after transplantation of anti-CD9 antibody-selected cells (2.7 3 104 cells injected). Note the increase in number of colonies of CD9-positive cells despite the lower concentration of injected cells. Bar 5 100 mm. C) Enhanced colonization of recipient testis by anti-CD9 antibody-selected rat testis cells. The number of individual fluorescent colonies represents the degree of colonization in three experiments. The values (mean 6 SEM) for anti-CD9 antibody-selected and unselected cells were 15.8 6 4.2 (n 5 16) and 3.1 6 0.9 (n 5 13) cells per 105 injected, respectively.

Article Snippet: Mouse anti-rat CD9 antibody was used with goat anti-mouse IgG microbeads (25 ml; Miltenyi Biotec) to select cells expressing rat CD9 molecules.

Techniques: Expressing, Cytometry, Staining, Control, Selection, Transplantation Assay, Injection, Concentration Assay

Journal: Molecular medicine reports

Article Title: Hypoxia-induced increases in A549/CDDP cell drug resistance are reversed by RNA interference of HIF-1α expression.

doi: 10.3892/mmr.2011.604

Figure Lengend Snippet: Figure 2. The effects of hypoxia and HIF-1α siRNA on the expression of MDR1 and MRP mRNAs in A549/CDDP cells. (A and C) The expression of MDR1 and MRP mRNA was measured by RT-PCR. M, DNA bp ladder; lane 1, control group; lane 2, hypoxia-treated group; lane 3, hypoxia- and control siRNA‑treated group; lane 4, hypoxia- and HIF-1α siRNA-treated-group. (B and D) Quantification of results shown in (A and C). MDR1, multidrug resis tance-1; MRP, multidrug resistance-associated protein; HIF-1α, hypoxia-inducible factor 1α; siRNA, small interfering RNA; RT-PCR, reverse transcription polymerase chain reaction.

Article Snippet: Antibodies to HIF-1α, MDR1 and MRP were purchased from Beijing Zhongshan Golden Bridge Biotechnology Company (China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Small Interfering RNA, Reverse Transcription, Polymerase Chain Reaction

Journal: Molecular medicine reports

Article Title: Hypoxia-induced increases in A549/CDDP cell drug resistance are reversed by RNA interference of HIF-1α expression.

doi: 10.3892/mmr.2011.604

Figure Lengend Snippet: Figure 5. The effects of hypoxia and HIF-1α siRNA on expression of MRP protein in A549/CDDP cells. (A-D) The expression of MRP protein was ana lyzed by immunocytochemistry. (A) Control group; (B) hypoxia-treated group; (C) hypoxia- and control siRNA-treated group; (D) hypoxia- and HIF-1α siRNA‑treated group. (E) Grading of positive protein staining in all 4 groups. HIF-1α, hypoxia-inducible factor 1α; siRNA, small interfering RNA; MRP, multidrug resistance‑associated protein.

Article Snippet: Antibodies to HIF-1α, MDR1 and MRP were purchased from Beijing Zhongshan Golden Bridge Biotechnology Company (China).

Techniques: Expressing, Immunocytochemistry, Control, Staining, Small Interfering RNA

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 6. Network-based analysis of interaction network corresponding to candidate genes. (A) The node degree distribution of PPI network. The number of genes is plotted as a function of their degree reflecting a power-law like distribution. The red line corresponds to a power-law distribution. (B) Sub-network of candidate genes. The large-sized black-colored nodes represent the candidate genes (ABCC1, ABCG2, CERK, MMP-2, MMP-9, and SPHK1), while the small gray color nodes represent the corresponding interactor genes. Interactions are represented in gray color and the depth of color represents the strength of correlations. (C) Shortest path lengths among candidate genes. Heatmap of shortest path length among the candidate genes, where the values represent the number of shortest paths between any pair.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 9. (A–D) Gene expression of drug transporters in breast cancer patients (A) ABCC1, (B) ABCG2 in local cohort (C) ABCC1, (D) ABCG2 in TCGA cohort and (E–G) Protein expression of drug transporters (E) Representative blots in adjacent normal (N) and tumor (T) tissues, (F) Densitometric analysis of ABCG2 and (G) ABCC1 levels in adjacent normal and tumor tissues.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques: Gene Expression, Expressing

Journal: Scientific reports

Article Title: Clinical relevance of CERK and SPHK1 in breast cancer and their association with metastasis and drug resistance.

doi: 10.1038/s41598-022-20976-0

Figure Lengend Snippet: Figure 10. Correlation of CERK with (A) ABCC1, (B) ABCG2 and SPHK1 with (C) ABCC1, (D) ABCG2 in local cohort and CERK with (E) ABCC1, (F) ABCG2 and SPHK1 with (G) ABCC1, (H) ABCG2 in TCGA cohort.

Article Snippet: Primary antibodies against SPHK1 (NBP2-20472); CERK (NB1002911); ABCC1 (NB400-156); ABCG2 (NBP2-22124); MMP-2 (NBP2-27208SS); MMP-9 (NBP2-13173SS) were purchased from Novus Biologicals (Centennial, Colorado, United States).

Techniques:

Journal: Pharmaceutics

Article Title: Targeted Self-Emulsifying Drug Delivery Systems to Restore Docetaxel Sensitivity in Resistant Tumors.

doi: 10.3390/pharmaceutics14020292

Figure Lengend Snippet: Figure 3. Enox-coated SEDDS inhibit MRP1 and BCRP activity. A459 cells were incubated for (a) 24 h or (c) 72 h with fresh medium (ctrl), 80 µM free docetaxel (DTX), 0.25% v/v blank SEDDS (S), Enox-coated SEDDS (S/Enox-Pa), SEDDs containing docetaxel (80 µM final concentration; S/DTX), and Enox-coated SEDDS containing docetaxel (80 µM final concentration; S/Enox-Pa/DTX). (a) The rate of ATP hydrolysis by immunopurified Pgp, MRP1, or BCRP extracted from cells treated as reported above was measured by spectrophotometric analysis in triplicates. Data are presented as means + SD (n = 3). * p < 0.02: vs. ctrl. (b) A549 cells were transduced with a non-targeting (scrambled) CRISPR-Cas vector or with a CRISPR-Cas vector to knock-out (KO) MRP1 or BCRP. The indicated proteins were measured by immunoblotting. Tubulin was used as control of equal protein loading. The image is representative of one out of three experiments. (c,d) The viability of scrambled, KO MRP1, and KO BCRP A549 cells was measured by a chemiluminescence-based assay in quadruplicates. Data are presented as means + SD (n = 3). * p < 0.001: vs. ctrl; ◦p < 0.001: S/Enox-Pa/DTX vs. DTX; # p < 0.01: S/Enox-Pa/DTX vs. S/DTX; § p < 0.001: KO vs. scrambled cells.

Article Snippet: The non-targeting siRNA sequence (Trilencer-27 Universal scrambled negative control siRNA duplex, #R30004), the FGFR1-targeting siRNAs pool of 3 unique 27mer siRNA duplexes (#SR320159), the CRISPR pCas vectors targeting MRP1 (#KN418182), BCRP (#KN405640), or the non-targeting vector (#GE100003) were purchased from Origene (Rockville, MD, USA).

Techniques: Activity Assay, Incubation, Concentration Assay, Transduction, CRISPR, Plasmid Preparation, Knock-Out, Western Blot, Control, Chemiluminescence Immunoassay