Journal: Cell Death & Disease

Article Title: Matrix stiffness regulates glucose-6-phosphate dehydrogenase expression to mediate sorafenib resistance in hepatocellular carcinoma through the ITGB1-PI3K/AKT pathway

doi: 10.1038/s41419-025-07842-3

Figure Lengend Snippet: A Venn diagram illustrating the overlap between genes related to ECM stiffness and those associated with sorafenib resistance. B Analysis of the top 20 KEGG pathways for genes related to ECM stiffness and sorafenib resistance. C Cross-validation plot for penalty terms. The cross-validation was used to select the optimum value of the regularization parameter ( λ ) in the LASSO regression model. D LASSO coefficient profiles of genes associated with ECM stiffness and sorafenib resistance. A coefficient profile plot was produced against the log( λ ) sequence. A vertical line was drawn at the selected optimal parameter ( λ ), which resulted in 12 nonzero coefficients. E A forest plot of 12 genes associated with ECM stiffness and sorafenib resistance. F Coefficients of these 12 genes. G Real-time qPCR analysis of gene expression related to ECM stiffness under soft (0.5 kPa) or stiff (12 kPa) matrix conditions. H Western blotting analysis of G6PD and AKR1B20 expression under soft (0.5 kPa) or stiff (12 kPa) matrix conditions. β-actin was used as a loading control and quantification data were shown on the right panel. I Representative images of H&E, Collagen I, α-SMA, Masson’s trichrome, Picrosirius red, AKR1B10, and G6PD staining in HCC tissues from patients who exhibited low stiffness and responsiveness to sorafenib and those with high stiffness and resistance to sorafenib. Scale bar, 200 and 500 μm as indicated. J Quantification of collagen I, α-SMA, Masson’s trichrome, and Picrosirius red area between 10 sorafenib-resistant samples with high ECM and 10 sorafenib-sensitive samples. K Association between AKR1B10 and G6PD expression levels and the degree of ECM stiffness in HCC tissue. ns not significance; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The primary antibodies used were as follows: G6PD (Proteintech, 25413–1-AP, 1:500), AKR1B10 (Proteintech, 18252–1-AP, 1:2000), ITGB1 (Proteintech, 12594–1-AP, 1:1000), collagen I (Proteintech, 10176–2-AP, 1:2500), α-SMA (Proteintech, 66009–1-Ig, 1:500), and p-AKT (Ser473, Proteintech, 66444–1-Ig, 1:2000).

Techniques: Biomarker Discovery, Produced, Sequencing, Gene Expression, Western Blot, Expressing, Control, Staining

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Primer sequences for real-time PCR amplification.

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Real-time Polymerase Chain Reaction, Amplification

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the mRNA expression levels of Zn, amino acid and small peptide transporters in the jejunum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the mRNA expression levels of Zn, amino acid and small peptide transporters in the ileum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the protein expression levels of Zn, amino acid and small peptide transporters in the jejunum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Representative immunoblots of ZnT1, ZnT4, ZnT5, ZnT7, ZnT9, ZIP3, ZIP5, B 0 AT1, LAT1, y+LAT2, rBAT and PepT1 in the jejunum of broilers at 28 (A) and 39 d (B) of age. ZnT1, zinc transporter 1; ZnT4, zinc transporter 4; ZnT5, zinc transporter 5; ZnT7, zinc transporter 7; ZnT9, zinc transporter 9; ZIP3, Zrt-irt-like protein 3; ZIP5, Zrt-irt-like protein 5; B 0 AT1, B-0-system neutral amino acid co-transporter; LAT1, L-type amino acid transporter 1; y+LAT2, y+L-type amino acid transporter 2; rBAT, b 0 , + -type amino acid transporter; PepT1, peptide-transporter 1; ZnS, Zn sulfate; Zn-Prot M, Zn proteinate with moderate chelation strength (Q f = 51.6).

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Western Blot

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Effect of dietary Zn source on the protein expression levels of Zn, amino acid and small peptide transporters in the ileum of broilers at 28 and 39 d of age. 1

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: The organic zinc with moderate chelation strength enhances the expression of related transporters in the jejunum and ileum of broilers

doi: 10.1016/j.psj.2023.102477

Figure Lengend Snippet: Representative immunoblots of ZnT1, ZnT4, ZnT5, ZnT7, ZnT9, ZIP3, ZIP5, B 0 AT1, LAT1, y+LAT2, rBAT and PepT1 in the ileum of broilers at 28 (A) and 39 d (B) of age. ZnT1, zinc transporter 1; ZnT4, zinc transporter 4; ZnT5, zinc transporter 5; ZnT7, zinc transporter 7; ZnT9, zinc transporter 9; ZIP3, Zrt-irt-like protein 3; ZIP5, Zrt-irt-like protein 5; B 0 AT1, B-0-system neutral amino acid co-transporter; LAT1, L-type amino acid transporter 1; y+LAT2, y+L-type amino acid transporter 2; rBAT, b 0 , + -type amino acid transporter; PepT1, peptide-transporter 1; ZnS, Zn sulfate; Zn-Prot M, Zn proteinate with moderate chelation strength (Q f = 51.6).

Article Snippet: The primary antibodies and dilution rates were as follows: ZnT1, ab214356, 1:1000 (Abcam, Cambridge, UK); ZnT4, BA3484, 1:1000 (Boster, Wuhan, China); Zn transporter 5 (ZnT5), 25604-1-AP, 1:1000 (Proteintech, Chicago); ZnT7, 13966-1-AP, 1:1000 (Proteintech, Chicago, IL); ZnT9, BS62531, 1:1000 (Bioworld, Minneapolis, MN); Zn transporter 10 ( ZnT10 ), PA5-49188, 1:1000 (Invitrogen, CA); ZIP3, ab254868, 1:500 (Abcam, Cambridge, UK); ZIP5, ab105194, 1:1000 (Abcam, Cambridge, UK); B 0 AT1, ab180516, 1:1000 (Abcam, Cambridge, UK); LAT1, DF8065, 1:500 (Affinity Biosciences, Melbourne, Australia); y+LAT2, 13823-1-AP, 1:1000 (Proteintech, Chicago); rBAT, DF7379, 1:1000 (Affinity Biosciences, Melbourne, Australia) and PepT1, A03672-1, 1:500 (Boster, Wuhan, China).

Techniques: Western Blot

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: Expression of AKR1B10 in CRC tissues. ( A ) Representative IHC images showing in situ AKR1B10 expression in CRC and normal tissues (scale bar = 100μm). ( B – E ) IHC scores of AKR1B10 in ( B ) CRC vs normal tissues, ( C ) T I-II vs T III-IV tissues, ( D ) tumors with or without lymph node invasion, and ( E ) early vs late TNM staging. ( F ) OS of AKR1B10 pos and AKRiB10 neg CRC patients in subgroups demarcated by tumor location, depth of tumor invasion, lymph node metastasis, degree of differentiation and TNM staging. ( G – I ) OS of ( G ) AKR1B10 pos and AKRiB10 neg CRC patients with TNM staging I-II ( H ) and III-IV ( I ). CRC, colorectal cancer. OS, overall survival. ns, no significant difference. ** P < 0.01, *** P < 0.001.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques: Expressing, In Situ

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: Relationship between AKR1B10 and clinic-pathological factors in 135 CRC patients.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques:

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: Results of univariate and multivariate analyses of postoperative patients’ survival by Cox’s proportional hazard model.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: Effect of AKR1B10 on CRC cell proliferation and migration ability. ( A ) Comparison of AKR1B10 mRNA expression in CRC and normal tissues across 7 Oncomine datasets. ( B – C ) AKR1B10 mRNA levels in ( B ) 27 paired CRC and normal tissues and ( C ) 5 CRC cell lines. ( D – E ) Immunoblots showing AKR1B10 protein levels in ( D ) wild type and ( E ) AKR1B10-KD and AKR1B10-OE CRC cell lines. ( F – H ) Proliferation rates ( F ), colony forming capacity ( G ) and migration rates ( H ) of AKR1B10-KD and AKR1B10-OE CRC cells. CRC, colorectal cancer. CTL, control; NC, negative control; KD, AKR1B10-shRNA; VEC, vector; OE, AKR1B10 overexpression plasmid. Data are presented as mean ± SD (n=3). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques: Migration, Comparison, Expressing, Western Blot, Control, Negative Control, shRNA, Plasmid Preparation, Over Expression

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: Correlation between AKR1B10 and FGF1 in CRC tissues. ( A ) Correlation analysis of AKR1B10 and FGF1 levels in CRC tissues from TCGA datasets by GEPIA platform. ( B ) FGF1 mRNA levels in 27 paired CRC and normal tissues. ( C ) Correlation between AKR1B10 and FGF1 levels in the above. ( D ) Representative IHC images showing in situ FGF1 expression in CRC and normal tissues (scale bar = 100μm) and ( E ) corresponding IHC scores. ( F ) OS of 135 CRC patients demarcated by FGF1 expression levels. ( G ) Stratification of 135 pairs of CRC and normal tissues into cluster 1 (red) and cluster 2 (green) according to AKR1B10 and FGF1 IHC scores. ( H ) Percentage of tumor and normal samples in each cluster. CRC, colorectal cancer. OS, overall survival. *** P < 0.001.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques: In Situ, Expressing

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: AKR1B10 knockdown suppresses CRC tumor growth in vivo . ( A – B ) Total body weight ( A ) and tumor volume ( B ) of the mice during the experiment. ( C ) Representative pictures of subcutaneous tumors harvested from NC and AKR1B10-KD group. ( D ) The weights of tumor masses. ( E ) Net body weight after subtracting the respective tumor weights. ( F – G ) Relative AKR1B10 ( F ) and FGF1 ( G ) mRNA levels in the tumors and their ( H ) correlation. ( I ) Stratification of mice into cluster 1 (grey) and cluster 2 (blue) according to body weight, tumor volume, tumor weight and AKR1B10 and FGF1 mRNA levels. ( J ) Percentage of NC and AKR1B10-KD mice in each cluster. Data are presented as mean ± SD. CRC, colorectal cancer. NC, negative control; KD, AKR1B10-shRNA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques: Knockdown, In Vivo, Negative Control, shRNA

Journal: Aging (Albany NY)

Article Title: Loss of AKR1B10 promotes colorectal cancer cells proliferation and migration via regulating FGF1-dependent pathway

doi: 10.18632/aging.103393

Figure Lengend Snippet: AKR1B10 inhibits CRC cell growth in an FGF1-dependent manner. ( A ) Immunoblot showing AKR1B10, FGF1 and GAPDH protein levels in HT29 cells transfected with AKR1B10-shRNA and in HCT116 cells transfected with AKR1B10 overexpression plasmid. ( B ) Immunoblot showing AKR1B10, FGF1 and GAPDH protein levels in HT29 transfected with FGF1-shRNA alone or in combination with AKR1B10-shRNA. ( C – E ) Proliferation rates ( C ), colony forming capacity ( D ) and migration rates ( E ) of the HT29 cells transfected as above. Data are presented as mean ± SD. NC, negative control; KD, AKR1B10-shRNA; VEC, vector; OE, AKR1B10 overexpression plasmid. “-”, control-shRNA. “+”, AKR1B10 or FGF1 shRNA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The processed sections were then blocked with 10% goat serum for 30 min, and incubated overnight with 1:200 diluted polyclonal anti-human AKR1B10 (BOSTER, Wuhan, China) or anti-human FGF1 (BOSTER, Wuhan, China) at 4°C.

Techniques: Western Blot, Transfection, shRNA, Over Expression, Plasmid Preparation, Migration, Negative Control, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Loss of the fructose transporter SLC2A5 inhibits cancer cell migration

doi: 10.3389/fcell.2022.896297

Figure Lengend Snippet: Attenuation of the SLC2A5 gene expression in fibrosarcoma cells. (A) . Images of scratch test (left) and quantitative analysis of closure (right) as a function of time for HT1080tdT and HT1080tdT-ε2A5 fibrosarcoma cells (n = 3). Scale bar 10 µm. (B) . Images of colonies formed by HT1080tdT and HT1080tdT-ε2A5 cells and analysis of colony number (500 cells/well plated at time 0 h). * p = 0.007; n = 3. Colonies were fixed with 70% ethanol and visualized by staining with 1% trypan blue. (C) . Proliferation assay of HT1080tdT and HT1080tdT-ε2A5 cells. HT1080tdT and HT1080tdT-ε2A5 cells were plated at a density of 5.0 × 10 3 cells/well of a 96 well plates. Fructose was added to the media as indicated in the Figure. Cells were cultured for 48 h followed by MTS assay (n = 3). All data in the Figure is representative of more than 3 experiments with 3 biological replicates.

Article Snippet: Anti-SLC2A5 antibodies were purchased from Boster (Cat# PA 2064) and used at 1:500 dilution.

Techniques: Gene Expression, Staining, Proliferation Assay, Cell Culture, MTS Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Loss of the fructose transporter SLC2A5 inhibits cancer cell migration

doi: 10.3389/fcell.2022.896297

Figure Lengend Snippet: Editing of the SLC2A5 gene. (A) . Inactivation of the SLC2A5 gene in human MIA-PaCa-2 and HT1080tdT cells using CRISPR/Cas9-mediated in situ gene editing. Guide RNAs were targeted to Exon 3. Genomic DNA of edited cells was sequenced, and positive clones were identified as having a 62 base pair disruption, leading to an early stop codon 14 amino acids downstream of the disruption point. (B) . qPCR analysis of SLC2A5 mRNA abundance in MIA-PaCa-2 and MIA-PaCa-ε2A5 cells (clones B3 and F11) (n = 3). DNA primers were designed to amplify Exon 12 of the SLC2A5 gene. Immunoblot analysis was carried out with antibodies directed at the C-terminus of SLC2A5. Anti-β-tubulin or anti-GAPDH antibodies were used to assess amounts of loaded protein samples. (C) . qPCR analysis of SLC2A5 mRNA abundance in HT1080tdT and HT1080tdT-ε2A5 cells. DNA primers and control antibodies were as described above. (D) . qPCR analyses of SLC2A1, SLC2A2, and SLC2A4 mRNA abundance in MIA-PaCa-2 and MIA-PaCa-ε2A5 cells (n = 3). (E) . qPCR analyses of SLC2A1, SLC2A2, and SLC2A4 mRNA abundance in HT1080tdT and HT1080tdT-ε2A5 cells (n = 3). All data in the Figure is representative of more than 3 biological replicates. NS, not significant.

Article Snippet: Anti-SLC2A5 antibodies were purchased from Boster (Cat# PA 2064) and used at 1:500 dilution.

Techniques: CRISPR, In Situ, Clone Assay, Disruption, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Loss of the fructose transporter SLC2A5 inhibits cancer cell migration

doi: 10.3389/fcell.2022.896297

Figure Lengend Snippet: Attenuation of the SLC2A5 gene expression in pancreatic cancer cells. (A) . Images of scratch test (left panel) and quantitative analysis of closure (right panel) for MIA-PaCa-2 and MIA-PaCa-ε2A5 cells (clone B3; ** p = 0.002, n = 3). Scale bar 10 µm. (B) . Scratch test analysis for MIA-PaCa-2 and MIA-PaCa-ε2A5 cell lines clone F11 (** p = 0.001, n = 3). Scale bar 10 µm. (C) . Colony formation images and colony number for MIA-PaCa-ε2A5 cells, clone B3 (left panel; 1000 cells/well plated at time 0 h; * p = 0.0001; n = 3) and clone F11 (right panel; 500 cells/well plated at time 0 h; * p = 0.001; n = 3). (D) . Fructose-dependent proliferation of the MIA-PaCa-2 and MIA-PaCa-ε2A5 cells. MIA-PaCa-2 and MIA-PaCa-ε2A5 (clones B3 and F11) cells were plated at a density of 1.0 × 10 4 cells/well of a 96 well plates. Fructose was added to the culture medium as indicated in the figure. The absorbance at 490 nm (OD 490 ) of the culture medium after MTS assay at time 0 and 48 h in culture are shown. Experiments were performed a minimum of three times in triplicates. * p = 0.0002; n = 3.

Article Snippet: Anti-SLC2A5 antibodies were purchased from Boster (Cat# PA 2064) and used at 1:500 dilution.

Techniques: Gene Expression, Clone Assay, MTS Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Loss of the fructose transporter SLC2A5 inhibits cancer cell migration

doi: 10.3389/fcell.2022.896297

Figure Lengend Snippet: Restoration of SLC2A5 function via SLC2A5 transgene expression. Proliferation assay (A) and scratch test assay (B) of MIA-PaCa-2, MIA-PaCa-ε2A5 (clone F11) and MIA-PaCa-ε2A5 cells expressing wild-type SLC2A5 or the non-functional SLC2A5-E401A mutant (* p = 0.002; n = 3). In (A) , fold change in cell number is shown at day 0 to day 3. In (B) , the percentage of open scratch is shown at time 0 and 48 h. Proliferation assay (C) and scratch test assay (D) of HT1080tdT, HT1080tdT-ε2A5 cells transfected with expression vectors encoding wild-type SLC2A5 or the non-functional SLC2A5-E401A mutant (* p = 0.001, n = 3). In (C) , the absorbance at 490 nm (OD 490 ) of the culture medium after MTS assay from time 0 and 7 h in culture is shown. In (D) , the percentage of open scratch is shown at time 0 and 7 h. NS, not significant. All data in the Figure is representative of more than 3 experiments with 3 replicates.

Article Snippet: Anti-SLC2A5 antibodies were purchased from Boster (Cat# PA 2064) and used at 1:500 dilution.

Techniques: Expressing, Proliferation Assay, Wound Healing Assay, Functional Assay, Mutagenesis, Transfection, MTS Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Loss of the fructose transporter SLC2A5 inhibits cancer cell migration

doi: 10.3389/fcell.2022.896297

Figure Lengend Snippet: Mitochondria localization and dynamics in SLC2A5-deficient cancer cells. (A) . Electron microscopy analysis of MIA-PaCa-2 cells shows that mitochondria were localized in the center of these cells in the perinuclear space (encircled). In MIA-PaCa-ε2A5 cells, mitochondria were spread throughout the cell towards the cell periphery, and appeared elongated (arrows) (n = 3). (B) . and (C) . Mitochondria area (B) and length (C) analyses in MIA-PaCa-2 and MIA-PaCa-ε2A5 cells. * p < 0.0006; ** p < 0.0001 (n = 3). (D) . Confocal images of mitochondria stained with MitoTracker ® Green in red fluorescent protein-labelled HT1080tdT cells,HT1080tdT-ε2A5 cells or HT1080tdT-ε2A5 cells expressing wild-type SLC2A5. Time-lapse video is shown in the (arrowhead). (E) . Mitochondria number and (F) . mitochondria length in fibrosarcoma HT1080tdT cells, HT1080tdT-ε2A5 or HT1080tdT-ε2A5 cells expressing wild-type SLC2A5. *** p < 0.0002 (n = 3).

Article Snippet: Anti-SLC2A5 antibodies were purchased from Boster (Cat# PA 2064) and used at 1:500 dilution.

Techniques: Electron Microscopy, Staining, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Loss of the fructose transporter SLC2A5 inhibits cancer cell migration

doi: 10.3389/fcell.2022.896297

Figure Lengend Snippet: SLC2A5 function is necessary for polarized localization of mitochondria in HT1080tdT human fibrosarcoma cells in vitro and in vivo . (A) mitochondria (green) localization in HT1080tdT or HT1080tdT-ε2A5 (vasculature, grey; blue, DAPI) cells that are extravasating out of the chicken CAM vasculature ( in vivo ). Right panels show all three channels: red fluorescent protein (red), mitochondria (green), nuclei (blue) and vasculature (grey). Time-lapse video is shown in the . (B) . Fluorescent channel intensity along the line scans (dashed arrows in (A) (right panel) indicate the direction of the scan). Green line depicts mitochondria localization. The short arrows in (A) and (B) indicate vascular membrane breaches.

Article Snippet: Anti-SLC2A5 antibodies were purchased from Boster (Cat# PA 2064) and used at 1:500 dilution.

Techniques: In Vitro, In Vivo, Membrane