Journal: International Journal of Medical Sciences

Article Title: Upregulation of C/EBP Homologous Protein induced by ER Stress Mediates Epithelial to Myofibroblast Transformation in ADTKD-UMOD

doi: 10.7150/ijms.65036

Figure Lengend Snippet: UMOD mutations in patients

Article Snippet: The primary antibodies used were: polyclonal sheep anti-human UMOD antibody (R&D Systems, USA; Cat #AF6144, 1:100), proximal tubule brush border labeling antibody LTL-488 (Fluorescein Lotus Lectin, Vector Laboratories, USA, FL-1321; 1:1000), anti-GRP78 BiP antibody (abcam, Cambridge, UK; ab21685; 1:500) and CHOP (L63F7) mouse mAb (Cell Signaling Technology, Danvers, MA, USA; #2895; 1:50).

Techniques: Mutagenesis, Variant Assay, Sequencing

Journal: International Journal of Medical Sciences

Article Title: Upregulation of C/EBP Homologous Protein induced by ER Stress Mediates Epithelial to Myofibroblast Transformation in ADTKD-UMOD

doi: 10.7150/ijms.65036

Figure Lengend Snippet: Clinical features of patients with UMOD mutations

Article Snippet: The primary antibodies used were: polyclonal sheep anti-human UMOD antibody (R&D Systems, USA; Cat #AF6144, 1:100), proximal tubule brush border labeling antibody LTL-488 (Fluorescein Lotus Lectin, Vector Laboratories, USA, FL-1321; 1:1000), anti-GRP78 BiP antibody (abcam, Cambridge, UK; ab21685; 1:500) and CHOP (L63F7) mouse mAb (Cell Signaling Technology, Danvers, MA, USA; #2895; 1:50).

Techniques: Biomarker Discovery, Immunofluorescence, Microscopy

Journal: International Journal of Medical Sciences

Article Title: Upregulation of C/EBP Homologous Protein induced by ER Stress Mediates Epithelial to Myofibroblast Transformation in ADTKD-UMOD

doi: 10.7150/ijms.65036

Figure Lengend Snippet: Representative histopathology findings of an ADTKD-UMOD renal biopsy (from CASE 2). (A, B) Hematoxylin-Eosin staining showing eosinophilic “fluffy” inclusions in thick ascending limb of Henle's loop (TALH) (arrow), i.e., abnormal protein accumulation. (C, D) Masson's trichrome staining showing interstitial fibrosis changes, obvious fibrosis around the distal tubules and an intracellular hyaline change (arrow). (E-G) Electron microscopy images; (E) a complete image of a distal tubular epithelium. (F, G) partial enlargement of (E) , showing that the rough ER and the smooth ER are obviously expanded, and accumulation of lower electron density in the ER, i.e., abnormal accumulation of protein.

Article Snippet: The primary antibodies used were: polyclonal sheep anti-human UMOD antibody (R&D Systems, USA; Cat #AF6144, 1:100), proximal tubule brush border labeling antibody LTL-488 (Fluorescein Lotus Lectin, Vector Laboratories, USA, FL-1321; 1:1000), anti-GRP78 BiP antibody (abcam, Cambridge, UK; ab21685; 1:500) and CHOP (L63F7) mouse mAb (Cell Signaling Technology, Danvers, MA, USA; #2895; 1:50).

Techniques: Histopathology, Staining, Electron Microscopy

Journal: International Journal of Medical Sciences

Article Title: Upregulation of C/EBP Homologous Protein induced by ER Stress Mediates Epithelial to Myofibroblast Transformation in ADTKD-UMOD

doi: 10.7150/ijms.65036

Figure Lengend Snippet: Uromodulin expression increases in kidneys of ADTKD-UMOD cases. Representative staining images of kidney paraffin sections from a healthy donor (HNK), sporadic chronic interstitial nephritis (CIN) case and two ADTKD-UMOD cases (CASE1, CASE2). Red, uromodulin; green, lotus tetragonolobus lectin (LTL; a marker of the brush border). Original magnification: 400×, scale bar, 20 µm; the areas in the white boxes were enlarged 4×. In HNK, uromodulin was mainly located on the cell membrane of distal tubular epithelial cells. In CIN, uromodulin was also distributed on the cell membrane, and the expression level did not significantly change compared with HNK. Secreted uromodulin protein was seen in the lumen in HNK and CIN (shown by arrows). In ADTKD-UMOD CASE1 and CASE2, there was significant uromodulin expression that aggregated in the cytoplasm of the distal tubular epithelial cells. Secreted uromodulin was not detected in ADTKD-UMOD.

Article Snippet: The primary antibodies used were: polyclonal sheep anti-human UMOD antibody (R&D Systems, USA; Cat #AF6144, 1:100), proximal tubule brush border labeling antibody LTL-488 (Fluorescein Lotus Lectin, Vector Laboratories, USA, FL-1321; 1:1000), anti-GRP78 BiP antibody (abcam, Cambridge, UK; ab21685; 1:500) and CHOP (L63F7) mouse mAb (Cell Signaling Technology, Danvers, MA, USA; #2895; 1:50).

Techniques: Expressing, Staining, Marker, Membrane

Journal: International Journal of Medical Sciences

Article Title: Upregulation of C/EBP Homologous Protein induced by ER Stress Mediates Epithelial to Myofibroblast Transformation in ADTKD-UMOD

doi: 10.7150/ijms.65036

Figure Lengend Snippet: Schematic diagram of CHOP promoting renal interstitial fibrosis in ADTKD-UMOD. Under normal physiological conditions, UMOD is synthesized and transported to the Golgi apparatus for processing, and then transported to the cell membrane. However, in ADTKD-UMOD, when mutant UMOD accumulates in the ER, it causes ER stress, which activates the ER membrane sensors. Consequently, GRP78 separates from the sensors and binds to the mutant UMOD to promote its folding; simultaneously the stress signal is transferred to the nucleus, resulting in high expression of CHOP and other ER chaperones (mainly GRP78). As a transcription regulator, CHOP promotes the expression of fibronectin and vimentin, which in turn cause the formation of renal interstitial fibrosis.

Article Snippet: The primary antibodies used were: polyclonal sheep anti-human UMOD antibody (R&D Systems, USA; Cat #AF6144, 1:100), proximal tubule brush border labeling antibody LTL-488 (Fluorescein Lotus Lectin, Vector Laboratories, USA, FL-1321; 1:1000), anti-GRP78 BiP antibody (abcam, Cambridge, UK; ab21685; 1:500) and CHOP (L63F7) mouse mAb (Cell Signaling Technology, Danvers, MA, USA; #2895; 1:50).

Techniques: Synthesized, Membrane, Mutagenesis, Expressing

Journal: The Journal of Cell Biology

Article Title: Extreme C-terminal sites are posttranslocationally glycosylated by the STT3B isoform of the OST

doi: 10.1083/jcb.201301031

Figure Lengend Snippet: Glycosylation of β-GUS in siRNA-treated cells. (A) Diagram of β-GUS showing the signal sequence (black), proteolytic cleavage sites (arrowheads), glycosylation sites, free cysteine residues (red diamonds), and a Myc-DDK tag. The four sequons are numbered 1–4; β-GUS mutants lacking one or more sequons are designated as β-GUSΔXYZ, in which XYZ is the list of mutated sequons. (B–D) Transfected HeLa cells were pulse labeled for 5 min and chased as indicated with (C, D, and F) or without (B) prior treatment with siRNAs. Anti-DDK immunoprecipitates were analyzed by SDS-PAGE to resolve β-GUS with zero to four N-linked glycans. (D) Pulse–chase analysis of glycosylation of the N 631 ET site in wild-type (WT) β-GUS (top) or β-GUSΔ123 (bottom). (E) The percentages of wild-type β-GUS chains that have four glycans (squares) or β-GUSΔ123 chains that have one glycan (circles and triangles) are shown. Triangles designate cells treated with siRNA for STT3B. Symbols in E are the mean of two determinations; error bars indicate individual data points. (F) Values shown below F are the mean of two determinations. (G) The kinetics ( t 1/2 values) of glycosylation of the N 631 ET site in the Δ12, Δ13, and Δ23 mutants was determined by pulse–chase analysis as in D and E. The values shown are the t 1/2 values obtained by fitting the experimental data to a single exponential equation; error bars designate standard deviations. Avg., average; EH, endoglycosidase H; NC, negative control.

Article Snippet: The Myc-DDK–tagged β-GUS, CD40 ligand, and CD69 expression vectors were purchased from OriGene.

Techniques: Sequencing, Transfection, Labeling, SDS Page, Pulse Chase, Negative Control

Journal: The Journal of Cell Biology

Article Title: Extreme C-terminal sites are posttranslocationally glycosylated by the STT3B isoform of the OST

doi: 10.1083/jcb.201301031

Figure Lengend Snippet: Glycosylation of Tf in siRNA-treated cells. (A) Diagram of transferrin (Tf) showing the signal sequence (black), glycosylation sites, disulfide bonds (red lines), NVT sequon insertion sites (I-1, I-2, and I-3), and C-terminal DDK-His tag. (B–H) Tf-DDK-His was immunoprecipitated with the anti-DDK antibody (B, C, and E–H), whereas untagged Tf was immunoprecipitated with the anti-Tf antibody (D). (B) HeLa cells transfected with DDK-His–tagged wild-type Tf or Tf single-site mutants (N432Q or N630Q) were pulse labeled for 5 min and chased as indicated. (C and D) Pulse–chase labeling (5-min pulse and 20-min chase) of DDK-His–tagged Tf (C) or untagged Tf (D) in siRNA-treated HeLa cells. Black line indicates that intervening lanes have been spliced out. (E–H) DDK-His–tagged Tf constructs that have NVT insertions were pulse labeled for 10 min with (F and G) or without (E and H) prior treatment of the cells with STT3B siRNA. (H) Immunoprecipitated samples were subjected to digestion with endoglycosidase H (EH) for brief periods as indicated to obtain digestion intermediates that were resolved by SDS-PAGE. Values listed below gel lanes (C and F) are the means of two determinations. (I) Murine glycopeptides derived from 1,902 proteins are plotted as a running sum (black squares) relative to distance from the C terminus. Linear regression fits (blue lines) of the two arms of the curve intersect at a length of 66 residues. Estimates of STT3B dependence of glycosylation of SHBG N 396 GT (black diamond), SHBG N 380 RS (gray diamond), Tf I-3 (open square), β-GUS-Myc-DDK N 631 ET (open diamond), Tf I-2ΔHis (gray square), Tf I-2 (gray circle), untagged Tf N 630 VT (open triangle), Tf I-1 (black square), Tf-DDK-His N 630 VT (gray triangle), CD40L-MycΔDDK (open circle), CD69-MycΔDDK (black circle), factor VII N 360 IT (inverted black triangle), and prosaposin N 429 ST (black triangle) are shown. Our scaling system is qualitative because STT3B depletions are incomplete. The STT3B dependence values are defined as follows based upon the observed reductions in sequon modification in STT3B-depleted cells: >50% reduction (+++, strong); 25–50% reduction (++, moderate); 5–25% reduction (+, detectable); and 0–5% reduction (−, independent). Values for factor VII N 360 IT and prosaposin N 429 ST are taken from . The red bar (OST-PT) designates the reported distance in residues between the OST active site and the ribosomal peptidyltransferase center. Avg., average; NC, negative control; WT, wild type.

Article Snippet: The Myc-DDK–tagged β-GUS, CD40 ligand, and CD69 expression vectors were purchased from OriGene.

Techniques: Sequencing, Immunoprecipitation, Transfection, Labeling, Pulse Chase, Construct, SDS Page, Derivative Assay, Modification, Negative Control

Journal: International Journal of Molecular Sciences

Article Title: Decreased Tissue Omega-6/Omega-3 Fatty Acid Ratio Prevents Chemotherapy-Induced Gastrointestinal Toxicity Associated with Alterations of Gut Microbiome

doi: 10.3390/ijms23105332

Figure Lengend Snippet: Decreased tissue n-6/n-3 ratio prevents CPT-11-induced alterations in the gut microbiome . ( A ) Principal coordinates analysis (PCOA) plot showing the results of Bray–Curtis distance-based analysis of beta diversity metrics. ( B ) Violin plot with lines at the median (dashed lines) and quartiles (complete lines) showing the differences in the Pielou’s evenness index. ( C ) Microbe–microbe interactions network [SparCC correlation analysis (WT+CPT-11 vs. FAT-1+CPT-11)]. Each node (*, GUSB-producing taxa; #, healthy gut making taxa) represents a taxon (colored based on the phylum level and sized based on the number of connections to that taxon). Two taxa are connected by an edge (co-occurrences: red; anti-occurrences: blue; p -value < 0.05 and correlation threshold 0.3; size reflects the magnitude). ( D ) Random Forests classification of taxa (genus level) in the vehicle (W and F1) or CPT-11 (WC and F1C) treated groups. ( E ) Phyla detected in the control and CPT-11 treated WT and FAT-1 mice. The numbers above each group show the relative abundance (RA) of the Proteobacteria phylum. ( F – K ) RA of differentially abundant (ANCOM test by QIIME2) bacterial groups such as Enterobacteriaceae ( F ) with representative colonic luminal contents MacConkey agar culture plate photos ( G ) showing the difference ( H ) in the growth of Escherichia Coli (pink colonies), Enterococcus ( I ), Bifidobacterium ( J ) and Akkermansia ( K ). ( L ) RA of beta-glucuronidase (GUSB)-producing bacteria measured using qPCR. ( M ) The difference in GUSB activity was measured at baseline (BL) and days (d) 6 using stool samples and at days 11 using cecal contents. ( N ) Immunohistochemical staining-based GUSB gene expression patterns in the proximal colon. ( O ) RA of GUSB (K01195) gene predicted using PICRUSt2. Data are shown as mean ± standard error of the mean. Data with different superscript letters are significantly different ( p < 0.05) according to the Kruskal–Wallis test ( B ) or Mann–Whitney test, or ordinary two-way ( M ) ANOVA followed by Sidak’s multiple comparisons test. Scale bar for images in ( J ) panel: 2000 μm.

Article Snippet: Paraffin blocks obtained from formalin-fixed colonic tissues ( n = 3 per control groups; n = 4 per CPT-11 treated groups) and primary antibodies for beta-glucuronidase (GUSB; 1:100; sc-374629), from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and for Myeloperoxidase [ ] (MPO; 1:100; GTX75318) from GeneTex (San Antonio, TX, USA) were given to Massachusetts General Hospital (MGH) Core, Boston, MA, USA.

Techniques: Control, Bacteria, Activity Assay, Immunohistochemical staining, Staining, Gene Expression, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Decreased Tissue Omega-6/Omega-3 Fatty Acid Ratio Prevents Chemotherapy-Induced Gastrointestinal Toxicity Associated with Alterations of Gut Microbiome

doi: 10.3390/ijms23105332

Figure Lengend Snippet: Dietary n-3 PUFA supplementation reduces CPT-11-induced alterations in the gut microbiome . ( A ) Principal coordinates analysis (PCOA) plot showing the results of Bray–Curtis distance-based analysis of beta diversity metrics. ( B ) Violin plot with lines at the median (dashed lines) and quartiles (complete lines) showing the differences in the Pielou’s evenness index (α-diversity). ( C ) Microbe–microbe interactions network [SparCC correlation analysis (CO+CPT-11 vs. FO+CPT-11)]. Each node (*, GUSB-producing taxa; #, healthy gut making taxa) represents a taxon (colored based on the phylum level and sized based on the number of connections to that taxon). Two taxa are connected by an edge (co-occurrences: red; anti-occurrences: blue; p -value < 0.05 and correlation threshold 0.3; size reflects the magnitude). ( D ) Random Forests classification of taxa (genus level) of CPT-11 treated CO and FO groups. ( E ) Composition summary showing the phyla detected in the CPT-11 treated CO and FO groups. The numbers above each bar show the relative abundance (RA) of the Proteobacteria phylum. ( F – I ) RA of differentially abundant bacterial groups such as Enterobacteriaceae ( F ) with representative colonic luminal contents MacConkey agar culture plate photos ( G ) showing the difference ( H ) in the growth of Escherichia Coli and Bifidobacterium ( I ), which is not detectable in CO group. ( J ) qPCR results showing the RA of beta-glucuronidase (GUSB)-producing bacteria. ( K ) The difference in GUSB activity was measured at baseline (BL) and days (d) 6 using stool samples and at days 11 using cecal contents. ( L ) Representative pictures are showing GUSB expression measured at proximal colon using the immunohistochemical technique. ( M ) RA of GUSB (K01195) gene predicted using PICRUSt2. Data are shown as mean ± standard error of the mean. Data with different superscript letters are significantly different ( p < 0.05) according to the nonparametric Mann–Whitney test (* p < 0.05, ** p < 0.01) or ordinary two-way ( K ) ANOVA followed by Sidak’s multiple comparisons tests. Scale bar for images in ( J ) panel: 2000 μm.

Article Snippet: Paraffin blocks obtained from formalin-fixed colonic tissues ( n = 3 per control groups; n = 4 per CPT-11 treated groups) and primary antibodies for beta-glucuronidase (GUSB; 1:100; sc-374629), from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and for Myeloperoxidase [ ] (MPO; 1:100; GTX75318) from GeneTex (San Antonio, TX, USA) were given to Massachusetts General Hospital (MGH) Core, Boston, MA, USA.

Techniques: Bacteria, Activity Assay, Expressing, Immunohistochemical staining, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Decreased Tissue Omega-6/Omega-3 Fatty Acid Ratio Prevents Chemotherapy-Induced Gastrointestinal Toxicity Associated with Alterations of Gut Microbiome

doi: 10.3390/ijms23105332

Figure Lengend Snippet: Decreased tissue n-6/n-3 ratio reduces CPT-11-induced imbalances in the host–gut microbiome interactions . ( A , B ) Host–microbiota interaction network built from Spearman’s nonparametric rank correlation coefficient ( p < 0.05) between host parameters and entire microbial parameters (genus-level) of WT+CPT-11 vs. FAT-1+CPT-1 ( A ) and CO+CPT-11 vs. FO+CPT-11 ( B ) comparisons. Nodes (filled squares) in panel A represent host parameters (cyan) and microbes (olive). Nodes in panel B represent host parameters (filled squares colored black) and microbes (different shapes indicate different phylum and colored light black). Lines (edges) represent statistically significant correlations ( p < 0.05) and are colored blue for positive and red for negative correlations. Edge size reflects the magnitude of the correlation. ( C ) Multiple Factor Analysis superimposing the host and gut microbiome (genus-level) data associated with a high tissue n-6/n-3 ratio (WT+CPT-11/CO+CPT-11 samples) and a low tissue n-6/n-3 PUFA ratio (FAT-1+CPT-11/FO+CPT-11 samples). Each line connects the host and microbial data from one sample. One end of each connecting line for an observation indicates the host (differently colored to indicate the groups), and another end (dark yellow) indicates the gut microbiota (GM). ( D ) Principal component analysis (PCA) of the host and gut microbiome (genus-level) data associated with a high n-6/n-3 ratio and a low n-6/n-3 PUFA ratio. ( E , F ) Biomarker analysis using multivariate [Random Forests (RF) classification with PLS-DA feature ranking method] receiver operator characteristic curve (ROC) based exploratory analysis performed on the combined host and microbial parameters. ( G ) Post-CPT-11 cecal contents beta-glucuronidase (GUSB) activity measurements were associated with a high n-6/n-3 ratio and a low n-6/n-3 PUFA ratio. ( H ) ROC curve generated with classical univariate ROC curve analysis showing the sensitivity and specificity for cecal contents GUSB activity. Data are shown as mean ± standard error of the mean. *** p < 0.001, nonparametric Mann–Whitney test.

Article Snippet: Paraffin blocks obtained from formalin-fixed colonic tissues ( n = 3 per control groups; n = 4 per CPT-11 treated groups) and primary antibodies for beta-glucuronidase (GUSB; 1:100; sc-374629), from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA) and for Myeloperoxidase [ ] (MPO; 1:100; GTX75318) from GeneTex (San Antonio, TX, USA) were given to Massachusetts General Hospital (MGH) Core, Boston, MA, USA.

Techniques: Biomarker Discovery, Activity Assay, Generated, MANN-WHITNEY

Journal: International Journal of Medical Sciences

Article Title: ABCC6 Transporter Contributed to Cisplatin Resistance on Bladder Cancer

doi: 10.7150/ijms.115487

Figure Lengend Snippet: Figure 1. Comparative Analysis of Cisplatin Resistance in T24 and UMUC3 BLCA Cells. (A&B) Cell viability of T24 and UMUC3 cells treated with varying concentrations of cisplatin (0 to 200 µM). (C&D) Colony formation assay for T24 and UMUC3 cells showing cisplatin resistance. T24R: T24 cell with cisplatin resistance; UMUC3R: UMUC3 cell with cisplatin resistance. *p> 0.05 vs. WT group.

Article Snippet: The cells were then treated with varying concentrations of cisplatin (0, 3.125, 6.25, 12.5, 25, 50, 100, 150, and 200 μM) for 24 h. Cell viability was evaluated using a resazurin reagent (Biotium, Inc.).

Techniques: Colony Assay

Journal: International Journal of Medical Sciences

Article Title: ABCC6 Transporter Contributed to Cisplatin Resistance on Bladder Cancer

doi: 10.7150/ijms.115487

Figure Lengend Snippet: Figure 2. The Role of ABC Transporters in Cisplatin Resistance of T24 and UMUC3 BLCA Cells. (A) mRNA expression levels of ABC transporters in T24 cells. (B) mRNA expression levels of ABC transporters in UMUC3 cells, (C) Protein levels of ABCC6 in T24 cells. (D) Protein levels of ABCC6 in UMUC3 cells. (E) Calcein AM efflux in T24 cells: (F) Calcein AM efflux in UMUC3 cells. * p <0.05, **p <0.01, *** p <0.001 and **** p <0.0001, vs. WT group.

Article Snippet: The cells were then treated with varying concentrations of cisplatin (0, 3.125, 6.25, 12.5, 25, 50, 100, 150, and 200 μM) for 24 h. Cell viability was evaluated using a resazurin reagent (Biotium, Inc.).

Techniques: Expressing

Journal: International Journal of Medical Sciences

Article Title: ABCC6 Transporter Contributed to Cisplatin Resistance on Bladder Cancer

doi: 10.7150/ijms.115487

Figure Lengend Snippet: Figure 3. The Role of ABCC6 in Cisplatin Resistance of T24 and UMUC3 BLCA Cells. (A) Western blot analysis showing ABCC6 protein levels in T24 cells under wild-type and cisplatin-resistant conditions. (B) Western blot analysis illustrating ABCC6 protein levels in UMUC3 cells under WT and cisplatin-resistant conditions. (C) Bar graph representing the fold changes in calcein AM efflux in T24 cells, comparing control and shABCC6-treated. (D) Bar graph illustrating the fold change in calcein AM efflux in UMUC3 cells, comparing control and shABCC6-treated conditions. * p <0.05, ** p <0.01, ***p<0.001 and **** p <0.0001, vs. control group.

Article Snippet: The cells were then treated with varying concentrations of cisplatin (0, 3.125, 6.25, 12.5, 25, 50, 100, 150, and 200 μM) for 24 h. Cell viability was evaluated using a resazurin reagent (Biotium, Inc.).

Techniques: Western Blot, Control

Journal: International Journal of Medical Sciences

Article Title: ABCC6 Transporter Contributed to Cisplatin Resistance on Bladder Cancer

doi: 10.7150/ijms.115487

Figure Lengend Snippet: Figure 4. Comparative Analysis of Autophagy and Protein Expression in Cisplatin-Resistant T24 and UMUC3 BLCA Cells. (A) Immunofluorescence staining of LC3-II and p62 in T24 cells (B) Western blot analysis of LC3 and p62 in T24 cells. (C) Western blot analysis of ATG5 and ATG12 in UMUC3 cells. (D) Western blot analysis of ATG5 and ATG12 in T24 cells. (E) AO staining in T24 cells. (F) AO staining in UMUC3 cells. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001, vs. WT group.

Article Snippet: The cells were then treated with varying concentrations of cisplatin (0, 3.125, 6.25, 12.5, 25, 50, 100, 150, and 200 μM) for 24 h. Cell viability was evaluated using a resazurin reagent (Biotium, Inc.).

Techniques: Expressing, Immunofluorescence, Staining, Western Blot

Journal: International Journal of Medical Sciences

Article Title: ABCC6 Transporter Contributed to Cisplatin Resistance on Bladder Cancer

doi: 10.7150/ijms.115487

Figure Lengend Snippet: Figure 5. The Role of ABCC6 in Drug Resistance of BLCA Cells. (A) ABCC6 mRNA expression in T24 cells treated with BafA1 (bafilomycin A1) or CQ (chloroquine). (B) ABCC6 mRNA expression in UMUC3 cells treated with BafA1 or CQ. (C) ABCC6 protein levels in T24 cells are treated with BafA1 or CQ. (D) ABCC6 protein levels in UMUC3 cells treated with BafA1 or CQ. (E) Calcein AM accumulation in T24 cells with cisplatin resistance treated with BafA1 or CQ. (F) Calcein AM accumulation in UMUC3 cells with cisplatin resistance treated with BafA1 or CQ. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001, vs. WT group.

Article Snippet: The cells were then treated with varying concentrations of cisplatin (0, 3.125, 6.25, 12.5, 25, 50, 100, 150, and 200 μM) for 24 h. Cell viability was evaluated using a resazurin reagent (Biotium, Inc.).

Techniques: Expressing

Journal: International Journal of Medical Sciences

Article Title: ABCC6 Transporter Contributed to Cisplatin Resistance on Bladder Cancer

doi: 10.7150/ijms.115487

Figure Lengend Snippet: Figure 6. Mechanism of Cisplatin Resistance Mediated by ABCC6 Transporters. This illustration highlights a crucial mechanism in chemotherapy resistance, emphasizing the role of ABCC6 transporters in reducing the intracellular concentration of cisplatin, thereby enhancing cell survival and resistance.

Article Snippet: The cells were then treated with varying concentrations of cisplatin (0, 3.125, 6.25, 12.5, 25, 50, 100, 150, and 200 μM) for 24 h. Cell viability was evaluated using a resazurin reagent (Biotium, Inc.).

Techniques: Concentration Assay