Journal: bioRxiv

Article Title: Replication-driven HBV cccDNA loss in chimeric mice with humanized livers

doi: 10.1101/2023.12.28.573542

Figure Lengend Snippet: A. Average cccDNA levels were <1 copy/cell in all 380 cccDNA samples isolated from livers of 17 mice under non-partial blocking of de novo infection (two mice: 970 and 819) or blocking of two cccDNA replenishment pathways (the remaining 15 mice). B. Average cccDNA levels were significantly lower than both untreated and treated mice with partial blocking of de novo infection . C1 and C2. Serum HBsAg and HBeAg levels were progressively reduced to undetectable levels among mice with non-partial blocking of de novo infection in mouse 819 or blocking of two pathways in the remaining 7 mice started before peak infection . C3. Kinetic serum anti-HBs antibody level . D1 and D2. Serum HBsAg and HBeAg levels were progressively reduced to undetectable levels upon boosting anti-HBs antibody levels by triweekly administrations of mouse anti-HBs antibody at a dose of 250 μg/injection starting on day 99 pi or later until termination. D3. Kinetic serum anti-HBs antibody level in the same D1/D2 group. E1 and E2. Average rcDNA levels reduced by 2-4 logs among mice through blocking of two pathways (Green) with anti-HBs antibody expressed by HBVZ10 before peak infection (E1) or with administering additional mouse anti-HBs antibody to boost the HBVZ10 expressed anti-HBs antibody level after the peak infection (E2) compared to untreated (Blue). F . Kinetic serum HBsAg levels among 3 mice (805 untreated; 833 HBVZ10 monotherapy, and 838 HBVZ10 combined with 12-week entecavir. G . Average intracellular HBsAg, rcDNA and cccDNA levels reduced by >100-fold in mouse 838 in 80 days compared to mouse 833. HBeAg, hepatitis B e antigen. The difference was considered significant at P<0.05 by Student’s t test. Error bars were plotted with standard deviations.

Article Snippet: Blood was collected tri-weekly for quantification of serum HBV DNA (qPCR see below), HBeAg (CSB-E13557h, CUSABIO), HBsAg (GS HBsAg EIA 32591, Bio-Rad), anti-HBs antibody (MONOLISA Anti-HBs EIA 25200, Bio-Rad) with calibrators (MONOLISA Anti-HBs 20-Calibrator kit 25219, Bio-Rad) and human albumin (Human albumin ELISA kit E-80AL, Immunology Consultants Laboratory) levels by ELISA per instructions.

Techniques: Isolation, Blocking Assay, Infection, Injection

Journal: Nature communications

Article Title: Syntenin-ALIX exosome biogenesis and budding into multivesicular bodies are controlled by ARF6 and PLD2.

doi: 10.1038/ncomms4477

Figure Lengend Snippet: Figure 5 | ARF6 depletion does not affect viral budding. (a) Western blot illustrating sustained ARF6 depletion during the time course of the experiment, viral release being measured 6 days after RNAi treatment. (b,c) MCF-7 cells infected with VSV-G-pseudotyped HIV-1 were analysed by confocal fluorescence microscopy to test for their ability to allow viral budding. (b) Viral envelope proteins were stained with anti-Env SUgp120 antibodies (green) and viruses that completed their budding were detected with an antibody recognizing mature matrix protein MAp17 (red). Note the high level of colocalization illustrating that most virions completed their budding at the plasma membrane. Scale bar, 5 mm. Nuclei were stained with DAPI (blue). (c) The total MAp17 fluorescence was divided by the total number of nuclei in each field, calculated for 150 to 200 cells in three independent experiments. Histograms represent the mean fluorescence intensity per nucleus±s.d. as an estimation of the number of mature viruses released per cell. NS, non significant (Student’s t-test). (d) After RNAi treatment, MCF-7 cells were infected with VSV-G-pseudotyped HIV-1. Three days after infection, cells were analysed by confocal fluorescence microscopy using a p24 antibody to stain intracellular Gag and the mature virus. The total p24 fluorescence was divided by the total number of nuclei in each field, calculated for 150 to 200 cells in three independent experiments. Histograms represent the mean fluore- scence intensity per nucleus±s.d. NS, non significant (Student’s t-test). (e) Virus release was determined by CAp24 quantification in the supernatant, using a HIV-1 p24 ELISA kit. p24 release was divided by the total number of cell. Results were normalized to 100% in siCNT cells. NS, non significant (Student’s t-test).

Article Snippet: Virus concentration was determined by CAp24 quantification in the supernatant, using a HIV-1 p24 ELISA kit (PerkinElmer Life Science, Inc.).

Techniques: Western Blot, Infection, Microscopy, Staining, Clinical Proteomics, Membrane, Virus, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Tumor-Derived Exosomes Induce the Formation of Neutrophil Extracellular Traps: Implications For The Establishment of Cancer-Associated Thrombosis

doi: 10.1038/s41598-017-06893-7

Figure Lengend Snippet: Exosome levels are increased in tumor-bearing mice. ( A ) Quantification of exosomes isolated from the plasma of control (●, n = 5) and 4T1 tumor-bearing (■, n = 10) mice. ( B ) Representative histogram indicating the size distribution of exosomes isolated from control (left panel) and 4T1 bearing-mice (right panel). Exosomes were isolated from the plasma as described in the Methods section. ( C ) CD63 was quantified in the plasma of control (open bar) and 4T1-bearing mice (black bar) as described in the Methods section. *P < 0.05; **P < 0.01; unpaired, two-tailed Student’s t-test.

Article Snippet: The mouse CD63 specific ELISA kit was from Cusabio (Wuhan, Hubei Province, China).

Techniques: Isolation, Clinical Proteomics, Control, Two Tailed Test

Journal: Pharmaceutics

Article Title: A Multivalent mRNA Therapeutic Vaccine Exhibits Breakthroughs in Immune Tolerance and Virological Suppression of HBV by Stably Presenting the Pre-S Antigen on the Cell Membrane

doi: 10.3390/pharmaceutics17020211

Figure Lengend Snippet: In vitro expression analysis of hepatitis B surface antigen mRNA vaccines. LNP-encapsulated L mRNA, M mRNA, and S mRNA were transfected into 293T cells in different combinations, and the dosage of each single mRNA was 5 μg for each treatment. ( a ) The proportion of cells expressing pre-S2 antigen or S antigen on the surface was detected by FCM at 48 h and 96 h after transfection, respectively. The results of FCM were showed using density maps. ( b ) WB analysis of the expression or secretion of the L protein (42 kDa), M protein (31 kDa), and S protein (27 kDa) in the cell membrane or supernatants at 48 h after transfection.

Article Snippet: An HBsAg ELISA Kit (E-EL-H6080, Elabscience Biotechnology Co., Ltd., Nanjing, China) was used to quantify the amount of HBsAg in the VLPs.

Techniques: In Vitro, Expressing, Vaccines, Transfection, Membrane

Journal: Pharmaceutics

Article Title: A Multivalent mRNA Therapeutic Vaccine Exhibits Breakthroughs in Immune Tolerance and Virological Suppression of HBV by Stably Presenting the Pre-S Antigen on the Cell Membrane

doi: 10.3390/pharmaceutics17020211

Figure Lengend Snippet: Humoral immune response analysis of HBV Tg mice immunized with the hepatitis B surface antigen mRNA vaccines. ( a ) Schematic diagram of the experimental protocol for the immunization of BALB/c HBV Tg mice with different combinations of mRNAs encoding hepatitis B surface antigens or with control vaccines. ( b ) CLIA detection of serum HBsAb levels in HBV Tg mice at weeks 0–32 of the immunization experiment. ( c ) Analysis of changes in the serum HBsAb concentration in mice whose HBsAb concentration was consistently less than 2000 IU/L and greater than 0 IU/L. ( d ) Correlation analysis between the serum levels of HBsAb and reduced HBsAg levels. ( e ) Indirect ELISA analysis of the binding ability of induced serum IgG to recombinant HBsAg, the pre-S peptide, or the LMS VLP after immunization with different vaccines. ( f ) Variable slope (four parameters) analysis via nonlinear regression (curve fit) was performed for the pre-S antigen-specific indirect ELISA. Representative results are presented as the means ± standard deviations (SDs).

Article Snippet: An HBsAg ELISA Kit (E-EL-H6080, Elabscience Biotechnology Co., Ltd., Nanjing, China) was used to quantify the amount of HBsAg in the VLPs.

Techniques: Vaccines, Control, Concentration Assay, Indirect ELISA, Binding Assay, Recombinant

Journal: Pharmaceutics

Article Title: A Multivalent mRNA Therapeutic Vaccine Exhibits Breakthroughs in Immune Tolerance and Virological Suppression of HBV by Stably Presenting the Pre-S Antigen on the Cell Membrane

doi: 10.3390/pharmaceutics17020211

Figure Lengend Snippet: Serological and virological response analysis of pAAV HBV1.2 mice immunized with hepatitis B surface antigen mRNA vaccines. ( a ) Schematic diagram of the experimental protocol for immunization of pAAV HBV1.2 mice with LMS mRNA vaccines, S mRNA, or GFP mRNA. Changes in serum ( b ) HBsAg, ( c ) HBsAb, and ( d ) HBeAg from 0 to 5 weeks after immunization of pAAV HBV-1.2 mice with different vaccines. ( e ) Q–PCR detection of serum HBV DNA levels in immunized mice at week 0 and week 5. Representative results are presented as the means ± SDs.

Article Snippet: An HBsAg ELISA Kit (E-EL-H6080, Elabscience Biotechnology Co., Ltd., Nanjing, China) was used to quantify the amount of HBsAg in the VLPs.

Techniques: Vaccines

Journal: Pharmaceutics

Article Title: A Multivalent mRNA Therapeutic Vaccine Exhibits Breakthroughs in Immune Tolerance and Virological Suppression of HBV by Stably Presenting the Pre-S Antigen on the Cell Membrane

doi: 10.3390/pharmaceutics17020211

Figure Lengend Snippet: Serological and virological response analysis of rAAV8 HBV1.3 mice immunized with hepatitis B surface antigen mRNA vaccines. ( a ) Schematic diagram of the experimental protocol for immunization of rAAV8 HBV1.3 (1 × 10 10 vg/mouse) mice with hepatitis B surface antigen mRNA vaccines in different combinations, S mRNA, or GFP mRNA. Changes in serum ( b ) HBsAg, ( c ) HBsAb, and ( d ) HBeAg from 0 to 9 weeks of immunization with different vaccines. ( e ) Q–PCR detection of serum HBV DNA levels in immunized mice at week 0 and week 9. Representative results are presented as the means ± SDs.

Article Snippet: An HBsAg ELISA Kit (E-EL-H6080, Elabscience Biotechnology Co., Ltd., Nanjing, China) was used to quantify the amount of HBsAg in the VLPs.

Techniques: Vaccines

Journal: Pharmaceutics

Article Title: A Multivalent mRNA Therapeutic Vaccine Exhibits Breakthroughs in Immune Tolerance and Virological Suppression of HBV by Stably Presenting the Pre-S Antigen on the Cell Membrane

doi: 10.3390/pharmaceutics17020211

Figure Lengend Snippet: The analysis of antigen-dependent T-cell responses activated by Hepatitis B surface antigen mRNA vaccines. ( a ) BALB/3T3 cells were transiently transfected with pD2531.L, pD2531.M, and pD2531.S plasmids (1:1:1 mass ratio) via Lipofectamine 3000, and FCM was performed to detect pre-S2 antigen and S antigen expression on the cell surface. ( b ) After immunization of BALB/c mice with different hepatitis B surface antigen mRNA vaccines or PBS, splenic CD4 + T cells were isolated and stimulated with BALB/3T3 cells transiently expressing the LMS antigen. Meanwhile, antigen-incubated T cells from PBS-immunized mice were used as negative control and CD3/CD28 bead-stimulated T cells from PBS-immunized mice were used as positive control. FCM was used to measure the level of IFN-γ in CD4 + T cells after the addition of brefeldin A.

Article Snippet: An HBsAg ELISA Kit (E-EL-H6080, Elabscience Biotechnology Co., Ltd., Nanjing, China) was used to quantify the amount of HBsAg in the VLPs.

Techniques: Vaccines, Transfection, Expressing, Isolation, Incubation, Negative Control, Positive Control

Journal: Pharmaceutics

Article Title: A Multivalent mRNA Therapeutic Vaccine Exhibits Breakthroughs in Immune Tolerance and Virological Suppression of HBV by Stably Presenting the Pre-S Antigen on the Cell Membrane

doi: 10.3390/pharmaceutics17020211

Figure Lengend Snippet: Schematic diagram of the mechanism by which the LMS mRNA therapeutic vaccine disrupts HBsAg-mediated immune tolerance and reactivates immune responses against HBV. The numbers 1–6 marked on the membrane structure of L protein represent intracellular structure of S antigen, T cell recognition epitopes of pre-S1 antigen, hepatocyte surface receptor NTCP binding site for pre-S1 antigen, S antigen (highly variable region) with a-antigen determinant, lipid membrane, hepatocyte binding region for pre-S2 antigen, membrane attachment region of HBV-infected hepatocytes, respectively.

Article Snippet: An HBsAg ELISA Kit (E-EL-H6080, Elabscience Biotechnology Co., Ltd., Nanjing, China) was used to quantify the amount of HBsAg in the VLPs.

Techniques: Membrane, Binding Assay, Infection

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Expression of EBV genes in PBMCs of Ts1–4. In Ts1 and Ts2, only EBNA1 and LMP1 were detected at week 4, respectively. In Ts3, BZLF1 was expressed from weeks 6 to 10, and LMP1 was observed only at week 6.In Ts4, BZLF1 was expressed from weeks 2 to 4, and the expression of LMP1 persisted throughout the observation period with the exception of week 4

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Expressing

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Expression of EBV genes in PBMCs of Ts7–10. In Ts7, the expression of LMP1 persisted throughout the observation period, excluding week 2, BZLF1 appeared at weeks 2 to 4,EA was detected intermittently from weeks 4 to 18, and EBNA1 was observed only at week 18. In Ts8, EBNA1, EBNA2 and LMP1were expressed at week 2. In Ts9, LMP1was detected at weeks 2, 6 and 21, and EA and EBNA1were detected only at weeks 2 and 16, respectively. In Ts10, the expression of BZLF1 persisted throughout the observation period,and LMP1 was detected at week 1

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Expressing

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Levels of EBV antibody in the serum of tree shrews post-infection. The level of VCA IgG increased to varying degrees in all tree shrews in which the EBV copy number increased; EBNA1 IgG increased in only1 tree shrew (Ts7),and EA IgG was not elevated ( a ). The levels of these antibodies were not increased in the EBV-infection-negative tree shrews ( b ) or in any of the 3 tree shrews that served as negative controls (data not shown)

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Infection

Journal: Virology Journal

Article Title: A study of Epstein-Barr virus infection in the Chinese tree shrew( Tupaia belangeri chinensis )

doi: 10.1186/s12985-017-0859-5

Figure Lengend Snippet: Western blotting for the detection of EBV gene expression in the livers and spleens. EBNA1 was detected in the spleens of Ts2, Ts3, Ts9 and Ts10. No EBNA1 expression was detected in the livers of any of the tree shrews. B95–8 cells and Ts11–13 were used as positive and negative controls, respectively. Ts5 and Ts6 were EBV-infection-negative. No EBNA1expression was detected inTs5, Ts6 or Ts11–13

Article Snippet: EBV antibodies in tree shrew serum were measured using an enzyme-linked immunosorbent assay (ELISA).The anti-EBV capsid antigen(VCA) IgG,EBV nuclear antigen (EBNA) IgG, and early antigen (EA) IgG were measured with the Epstein-Barr virus VCA IgG ELISA Kit(Abnova,Taiwan, China),a human Epstein-Barr virus nuclear antigen (EBNA1) antibody (IgG) ELISA kit (CUSABIO, Wuhan, China), and a human Epstein-Barr virus early antigen (EBEA) antibody (IgG) ELISA kit (CUSABIO).Tree shrew serum diluted 1:10 with dilution buffer was placed into each plate, and bound antibodies were subsequently detected using rabbit anti-tree shrew IgGsecondary antibody synthesized by Sangon Biotech Co.,Ltd. (Shanghai, China).

Techniques: Western Blot, Gene Expression, Expressing, Infection

Journal: Molecular Oncology

Article Title: Extracellular LGALS3BP : a potential disease marker and actionable target for antibody–drug conjugate therapy in glioblastoma

doi: 10.1002/1878-0261.13453

Figure Lengend Snippet: LGALS3BP expression levels in GBM patients. (A) Sandwich ELISA performed on GBM patient ( n = 17) and healthy donor ( n = 18) serum samples. (B) Graph showing the percentage of EVs‐associated LGALS3BP over total circulating LGALS3BP in serum. Mann–Whitney test * P = 0.03. (C) Box‐and‐whisker diagram of the distribution of LGALS3BP in GBM cases and peritumoral matched samples ( n = 53). The upper and lower ends of boxes represent 75th and 25th percentiles. The median value is shown with a solid line. Mann–Whitney test *** P < 0.001. Data are shown as mean ± standard deviation. (D, E) Representative images of IHC staining for LGALS3BP expression in GBM and peritumoral matched samples. All IHC images have been acquired using an optical microscope at a scale bar of 50 μm.

Article Snippet: Also, for LGALS3BP expression analysis of mice serum‐derived EVs in correlation studies, they were analysed using Proteintech™ Human LGALS3BP ELISA Kit (Cod.

Techniques: Expressing, Sandwich ELISA, MANN-WHITNEY, Whisker Assay, Standard Deviation, Immunohistochemistry, Microscopy

Journal: Molecular Oncology

Article Title: Extracellular LGALS3BP : a potential disease marker and actionable target for antibody–drug conjugate therapy in glioblastoma

doi: 10.1002/1878-0261.13453

Figure Lengend Snippet: LGALS3BP expression levels in EVs isolated from GBM patient‐derived cell lines. (A) LGALS3BP and EVs markers CD9, actin and CD63 expression levels in whole lysate and EVs isolated from Gch6 and Gch14 cellular supernatants ( n = 2). (B) Effect of N‐Glycosylation (kifunensine, KIF and tunicamycin, TUN) and O‐Glycosylation (OSMI‐1) inhibitors on the protein profile for LGALS3BP in Gch6 whole lysate and corresponding supernatant ( n = 2). (C) Sandwich ELISA performed on intact EVs isolated from Gch6 and Gch14 cell line supernatants ( n = 3). Data are shown as mean ± standard deviation. (D) Sandwich ELISA performed on intact EVs isolated from Gch6 treated with KIF ( n = 3). Data are shown as mean ± standard deviation. (E) Effect of KIF treatment on the protein profile in whole lysate and EVs isolated from Gch6 ( n = 2). (F) Confocal images of live GBM patient‐derived cells labelled with humanized 1959 anti‐LGALS3BP antibody followed by AlexaFluor 488 conjugated secondary anti‐human IgG antibody (green). Cell nuclei were stained with DAPI (blue) ( n = 3). Images were taken at 40× magnification. Scale bar: 20 μm. Negative controls were only incubated with secondary antibody.

Article Snippet: Also, for LGALS3BP expression analysis of mice serum‐derived EVs in correlation studies, they were analysed using Proteintech™ Human LGALS3BP ELISA Kit (Cod.

Techniques: Expressing, Isolation, Derivative Assay, Sandwich ELISA, Standard Deviation, Staining, Incubation

Journal: Molecular Oncology

Article Title: Extracellular LGALS3BP : a potential disease marker and actionable target for antibody–drug conjugate therapy in glioblastoma

doi: 10.1002/1878-0261.13453

Figure Lengend Snippet: Extracellular vesicles‐associated LGALS3BP correlation with tumour volume and therapeutic activity of 1959‐sss/DM3 in GBM patient‐derived xenograft model. (A) Graphic scheme representing ELISA of human LGALS3BP on EVs in serum samples from mice with human GBM xenograft. (B) Levels of LGALS3BP on EVs isolated from the serum samples of control nude mice or mice bearing human GCh6 GBM xenograft, measured by ELISA ( n = 5). Bar graph represents average ± standard deviation. (C) Spearman correlation between EVs‐associated LGALS3BP and tumour burden in xenograft‐bearing nude mice ( n = 9). (D, E) Cytotoxic activity of SH‐DM3, SH‐DM4 and temozolomide (TMZ) was obtained by using the MTT assay on Gch6 and Gch14 cell lines at indicated doses after 5 days. IC50 values were determined with graphpad prism software (San Diego, CA, USA) ( n = 3). Data are shown as mean ± standard deviation. (F) CD1 nude mice harbouring Gch6 xenografts were treated with vehicle (PBS, n = 4) or 1959‐sss/DM3 (10 mg·kg −1 , n = 4) twice weekly as indicated by the arrows. Data are shown as mean ± standard error. (G) (PBS, n = 4) (10 mg·kg −1 , n = 4). Kaplan–Meier survival curves. Log‐rank (Mantel–Cox) Test. ** P = 0.006.

Article Snippet: Also, for LGALS3BP expression analysis of mice serum‐derived EVs in correlation studies, they were analysed using Proteintech™ Human LGALS3BP ELISA Kit (Cod.

Techniques: Activity Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Isolation, Standard Deviation, MTT Assay, Software

Journal: Molecular Oncology

Article Title: Extracellular LGALS3BP : a potential disease marker and actionable target for antibody–drug conjugate therapy in glioblastoma

doi: 10.1002/1878-0261.13453

Figure Lengend Snippet: 1959‐sss/DM4 dose–response efficacy and biodistribution. (A) CD1 nude mice Gch6 xenografts were treated with vehicle (PBS) or 1959‐sss/DM4 at the indicated doses twice weekly (PBS, n = 5; 1 mg·kg −1 , n = 7; 3 mg·kg −1 , n = 6; 10 mg·kg −1 , n = 6). Data are shown as mean ± standard error. (B) (PBS, n = 5; 1 mg·kg −1 , n = 7; 3 mg·kg −1 , n = 6; 10 mg·kg −1 , n = 6). Kaplan–Meier survival curves. Log‐rank (Mantel–Cox) Test. *** P < 0.001. (C) Representative images of IHC staining for LGALS3BP expression in Gch6 patient primary tumour and in Gch6 xenograft ( n = 4). Scale bar: 50 μm. (D) Biodistribution of 1959‐sss/DM4 in tumour‐bearing nude mice. ADC was injected into the tail vein of nude mice bearing a xenograft of GCh6 patient‐derived cell line. After 24, 48 and 72 h, the mice were sacrificed, and the ADC uptake was determined in tissues (D) by ELISA using an anti‐DM4 antibody as coating. Each column and bar show the mean and standard deviation for three mice. Tissue uptake of ADC is expressed as percentage of injected dose per gram of tissue (%ID·g −1 ). Data are shown as mean ± standard deviation.

Article Snippet: Also, for LGALS3BP expression analysis of mice serum‐derived EVs in correlation studies, they were analysed using Proteintech™ Human LGALS3BP ELISA Kit (Cod.

Techniques: Immunohistochemistry, Expressing, Injection, Derivative Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation