Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Primer sequences of OCT2, OAT1, OAT3, P-gp, MATE1, MATE2K, MRP2 genes.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Sequencing

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Values of the binding energies of berberine and berberrubine to seven transporter proteins in the kidney (Kcal/mol).

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Binding Assay

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Molecular docking illustrations of BBR and BBRR with seven transport proteins respectively. (A) BBR docked with OAT1. (B) BBR docked with OAT3. (C) BBR docked with OCT2. (D) BBR docked with MATE1. (E) BBR docked with MATE2K. (F) BBR docked with P‐gp. (G) BBR docked with MRP2. (H) BBRR docked with OAT1. (I) BBRR docked with OAT3. (J) BBRR docked with OCT2. (K) BBRR docked with MATE1. (L) BBRR docked with MATE2K. (M) BBRR docked with P‐gp. (N) BBRR docked with MRP2.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: Trends of metformin, methotrexate, estrone-3-sulfate, topotecan, rhodamine 123, and vinblastine concentration under different dosing time conditions in cells lysates or extracellular medium from HEK-293 cells. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR). (A) The trend of MET concentration changes mediated by OCT2. (B) The trend of MTX concentration changes mediated by OAT1. (C) The trend of E3S concentration changes mediated by OAT3. (D) The trend of Rho123 concentration changes mediated by P-gp. (E) The trend of TOP concentration changes mediated by MATE1 and MATE2K. (F) The trend of VBL concentration changes mediated by MRP2 (n = 3).

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Concentration Assay, Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: The effect of various solutions on each substrate accumulation transported by renal transport proteins. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR) (A) The effect of various solutions on the accumulation of MET transported by OCT2. (B) The effect of various solutions on the accumulation of MTX transported by OAT1. (C) The effect of various solutions on the accumulation of E3S transported by OAT3. (D) The effect of various solutions on the accumulation of Rho123 transported by P-gp. (E) The effect of various solutions on the accumulation of TOP transported by MATE1 and MATE2K. (F) The effect of various solutions on the accumulation of VBL transported by MRP2. Compared with the RPC group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; compared with the SPC group, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 (n = 3).

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: The effect of various solutions on the protein expression of MATE1, MATE2K, P-gp, MRP2. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR). (A) MATE1, MATE2K, P-gp, MRP2 protein expression bands in HEK-293 cells. (B) The effect of various solutions on MATE1 protein expression level. (C) The effect of various solutions on MATE2K protein expression level. (D) The effect of various solutions on P-gp protein expression level. (E) The effect of various solutions on MRP2 protein expression level. Compared with the BC group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; compared with the RPC group, □ p < 0.05, □□ p < 0.01, □□□ p < 0.001, □□□□ p < 0.0001; compared with the BBR group, △ p < 0.05, △△ p < 0.01, △△△ p < 0.001, △△△△ p < 0.0001.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: A strategy for evaluating the impact of processing of Chinese meteria medica on meridian tropism: the influence of salt-water processing of phellodendri chinensis cortex on renal transport proteins

doi: 10.3389/fphar.2025.1558298

Figure Lengend Snippet: The effect of various solutions to MATE1, MATE2K, P-gp, MRP2 mRNA expression level. Blank control (BC), raw Phellodendri Chinensis cortex (RPC), Phellodendri Chinensis Cortex with salt-water processing (SPC), berberine (BBR), berberrubine (BBRR). (A) The effect of various solutions on MATE1 mRNA expression level. (B) The effect of various solutions on MATE2K mRNA expression level. (C) The effect of various solutions on P-gp mRNA expression level. (D) The effect of various solutions on MRP2 mRNA expression level Compared with the BC group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; compared with the RPC group, □ p < 0.05, □□ p < 0.01, □□□ p < 0.001, □□□□ p < 0.0001; compared with the BBR group, △ p < 0.05, △△ p < 0.01, △△△ p < 0.001, △△△△ p < 0.0001.

Article Snippet: DMEM high glucose medium (Cat. No.: 8123207), PBS buffer salt solution (Cat. No.: 20210927) were purchased from Gibco (Waltham, Massachusetts, United States), fetal bovine serum (Cat. No.: ST200913) was purchased from PAN (Aden Bach, Free State of Bavaria, Germany), penicillin-streptomycin-Amphotericin B Mixed Triple Antibody Solution (Cat. No.: 20220519JH), 20×TBST buffer (Cat. No.: T1082), Whole Protein Lysis Kit (Cat. No.: BC3710), TRIzol Lysis Solution (Cat. No.: 15596018CN), ECL PLUS Ultra-Sensitive Luminescent Solution (Cat. No.: PE0010), dimethyl sulfoxide (Cat. No.: 710N0310), metformin (Cat. No.: D9351, purity≥98%), methotrexate (Cat. No.: M8971, purity≥98%), rhodamine123 (Cat. No.: R8030, purity≥98%), topotecan (Cat. No.: IT1030, purity≥98%) were purchased from Solarbio Science and Technology Co. Ltd (Beijing, China), SDS-PAGE Protein Sampling Buffer 5× (Cat. No.: P0015L) was purchased from Beyotime Bio-Tech Ltd (Shanghai, China), OCT2 rabbit anti-polyclonal antibody (Cat. No.: 10867-2-AP), OAT1 rabbit anti-polyclonal antibody (Cat. No.: 26574-1-AP), OAT3 rabbit anti-polyclonal antibody (Cat. No.: 16844-1-AP), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20,898), P-gp rabbit anti-polyclonal antibody (Cat. No.: 22336-1-AP), MATE1 rabbit anti-polyclonal antibody (Cat. No.: 20898-1-AP), MATE2K rabbit anti-polyclonal antibody (Cat. No.: 26873-1-AP), MRP2 rabbit anti-polyclonal antibody (Cat. No.: 29261-1-AP), Goat Anti-Rabbit IgG II Antibody (Cat. No.: 20001097) were purchased from the Proteintech (Wuhan, China).

Techniques: Expressing, Control

Journal: Circulation

Article Title: Proteomic Analysis of Lung Tissues From Patients With Pulmonary Arterial Hypertension

doi: 10.1161/circulationaha.110.972745

Figure Lengend Snippet: Figure 4. Western blot analysis of lung homogenates. Equal amounts of protein (20 g) were used for Western blot analysis of PAH (n14) and control (n14) lung homogenates, using antibodies against periostin, CLIC1, CLIC4, haptoglobin, and vinculin. The densi- tometry results (arbitrary units [AU]) of the Western blots are shown as a bar graph. Each bar represents meanSEM. Representative Western blots for each protein are also shown. *P0.05, **P0.01, ***P0.001 vs control.

Article Snippet: Sections were immunostained with an affinity purified rabbit antibody against a recombinant protein epitope (73-217 amino acids) of human CLIC4 (HPA008019; Atlas Antibodies, Stockholm, Sweden; 1-2 μg/ml) and a recombinant human periostin fusion protein (ab14041, Abcam plc; 5 μg/ml).

Techniques: Western Blot, Control

Journal: Circulation

Article Title: Proteomic Analysis of Lung Tissues From Patients With Pulmonary Arterial Hypertension

doi: 10.1161/circulationaha.110.972745

Figure Lengend Snippet: Figure 6. Distribution of CLIC4 immuno- staining in human lung. Sections from lobectomy (A and B) and unused donor lung tissues (C), showing CLIC4 local- ized to bronchiolar epithelium (A), alveo- lar type II cells (open arrows), and mac- rophages (B), and endothelium (arrows) in the pulmonary vasculature (C). Sec- tions of lung from patients with IPAH (D through F), showing CLIC4 and CD31 immunostaining of the endothelium (arrows) in remodeled pulmonary arteries with neointimal proliferation, as well as the distal microvasculature (arrowheads). Cells in the neointimal layer display weaker CLIC4 immunostaining (D and F). Bar50 m.

Article Snippet: Sections were immunostained with an affinity purified rabbit antibody against a recombinant protein epitope (73-217 amino acids) of human CLIC4 (HPA008019; Atlas Antibodies, Stockholm, Sweden; 1-2 μg/ml) and a recombinant human periostin fusion protein (ab14041, Abcam plc; 5 μg/ml).

Techniques: Immunostaining

Journal: Circulation

Article Title: Proteomic Analysis of Lung Tissues From Patients With Pulmonary Arterial Hypertension

doi: 10.1161/circulationaha.110.972745

Figure Lengend Snippet: Figure 7. Distribution of CLIC4 immuno- staining in a plexiform lesion. Adjacent sections of a plexiform lesion, showing the localization of CLIC4 and CD31 in the endothelium (arrows) and CLIC4 in underlying smooth muscle cells or myo- fibroblasts (asterisks). Boxed area (A) shown at higher power (C). Bar50 m.

Article Snippet: Sections were immunostained with an affinity purified rabbit antibody against a recombinant protein epitope (73-217 amino acids) of human CLIC4 (HPA008019; Atlas Antibodies, Stockholm, Sweden; 1-2 μg/ml) and a recombinant human periostin fusion protein (ab14041, Abcam plc; 5 μg/ml).

Techniques: Immunostaining

Journal: Mechanisms of Development

Article Title: The zebrafish goosepimples/myosin Vb mutant exhibits cellular attributes of human microvillus inclusion disease ☆ ☆☆

doi: 10.1016/j.mod.2016.08.001

Figure Lengend Snippet: The loss of intestinal folds in gsp/myoVb mutants is accompanied by changes in cell shapes. A schematic showing various regions of the zebrafish larval gut (A). The boxed regions indicate the parts of the intestine where whole mounts were imaged (I) or intestines were sectioned (II, III). Confocal images of the whole mount (B, F), orthogonal sections (C, G), immuno-histological sections (D, E, H, I, J, K) of wild type sibling (B, C, D, E, J) and gsp/myoVb mutant (F, G, H, I, K) intestines stained for lyn-EGFP at 6dpf. Note the absence of intestinal folds in the gsp/myoVb mutants. The whole mount analysis was done on 4 siblings and 3 mutants whereas the immunohistology analysis was done on 9 siblings and 7 mutants. Bar graph (L) showing measurements of the height, apical width and basal width of enterocytes in wild type and gsp/myoVb mutant larvae at 6dpf. The enterocytes are taller and have a narrower apical domain in the gsp/myoVb mutants as compared to wild type. BrdU staining in wild type (M) and gsp/myoVb mutant larvae (N) at 3dpf followed by estimation of proliferation indices, which are represented in a bar graph (O). Number of enterocytes per 100 μm gut perimeter length was estimated from sections for 6-day-old wild type and gsp/myoVb mutant larvae and shown in a bar graph (P). Immuno-histological sections of zebrafish intestine in the wild type (Q) and gsp/myoVb mutant larvae (R) stained using anti E-cadherin and anti-Lgl2 antibodies. Arrows in B–E and J point to the intestinal folds. The square brackets show the comparison and asterisks indicate the statistically significant difference at p < 0.05 by Student's t -test. The error bars in L, O, and P represent SEM. Scale bars are equivalent to 20 μm. Abbreviation: sib = sibling.

Article Snippet: These sections were blocked with 10% NGS for 3 h. The whole intestinal tissue and cryosections were incubated overnight in anti-E-cadherin antibody (1:100; BD Transduction Labs, USA; #610182), anti-GFP antibody (1:200; Torrey Pines Biolabs, USA; #TP401), anti-Lgl2 antibody (1:400; , anti-Na + K + ATPase (1:100; DSHB; #a5, deposited by Fambrough, D.M.), anti-pEzrin (T567)/Radixin (T564)/Moesin (T558) (1:50; Cell signalling technology; #3141S), anti-pan Cytokeratin AE1/AE3 (1:100; Abcam; ab27988), anti-BrdU (1:50; Acris antibodies, #SM1667PS), Alexa Fluor 594 conjugated WGA (5 μg/ml; Invitrogen, USA; #W11262), Rhodamine Phalloidin (1:40; Invitrogen, USA, #R415) or Atto 647N-Phalloidin (1:400; Sigma, USA; #65906) in 1% NGS in PBT.

Techniques: Mutagenesis, Staining, BrdU Staining

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) The expression of HIF-1α and HIF-2α in gastrointestinal vascular malformations and normal vessels. Red arrow: strongly positive; Black arrow: weakly positive; Blue arrow: negative. ( B ) Percentages of positive and negative vessels in GIVM and normal tissues. ** P < 0.01.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) Western blot determinations of HIF-1α and HIF-2α expression at different time points of hypoxia. * P < 0.05, ** P < 0.01 vs. 0 hour. ( B ) The effect of HIF-1α and HIF-2α overexpression on the expression of VEGF, Notch1, DLL4, and Ang2. Western blot and RT-PCR demonstrated that HIF-1α and HIF-2α overexpression increased the expression of VEGF, Notch1, DLL4, and Ang2 protein and mRNA. * P < 0.05, ** P < 0.01 vs. control. ( C ) Influence of HIF-1α and HIF-2α overexpression on angiogenesis 6 and 24 h after transfection of Lenti-HIF-1α and Lenti-HIF2α. Tube formation was enhanced 6 and 24 h after transfection. ** P < 0.01 vs. control. ( D ) Fluorescence microscope observations of subintestinal vein sprouting in normal and HIF-2α-overexpressing zebrafish. *Indicates subintestinal vascular sprouts. HIF-2α overexpression significantly increased the number of subintestinal vascular sprouts. ** P < 0.01 vs. control plasmid. ( E ) Dual luciferase reporter gene assay demonstrated that HIF-2α enhanced VEGF promoter activity. ** P < 0.01 vs. control plasmid.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Western Blot, Expressing, Over Expression, Reverse Transcription Polymerase Chain Reaction, Transfection, Fluorescence, Microscopy, Plasmid Preparation, Luciferase, Reporter Gene Assay, Activity Assay

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) Immunofluorescence indicated that HIF-1α and HIF-2α expression was down-regulated by thalidomide at different concentrations. ( B ) Western blots demonstrated that the expression of HIF-1α and HIF-2α decreased with 100 and 200 μg/ml of thalidomide. * P < 0.05, ** P < 0.01. ( C ) Western blots demonstrated that thalidomide at 200 μg/ml inhibited the expression of HIF-1α and HIF-2α in HUVECs after hypoxic treatment for 24, 36, and 48 h. * P < 0.05, ** P < 0.01. ( D ) Fluorescence microscope observations of the effect of thalidomide at different concentrations on vascular development in zebrafish with HIF-2α overexpression. ** P < 0.01 vs. HIF-2α.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Immunofluorescence, Expressing, Western Blot, Fluorescence, Microscopy, Over Expression

Journal: Journal of Biological Chemistry

Article Title: B-Raf Contributes to Sustained Extracellular Signal-regulated Kinase Activation Associated with Interleukin-2 Production Stimulated through the T Cell Receptor

doi: 10.1074/jbc.m403087200

Figure Lengend Snippet: FIG. 3. Ras-regulated B-Raf activation following TCR stimula- tion in Jurkat cells. A, TAg-Jurkat cells were transfected with a mock vector or with the RasN17 expression vector, and then these cells were harvested and stimulated with cross-linking of soluble anti-CD3 anti- body for the indicated times. Immunoprecipitates from each cell extract with an anti-B-Raf antibody were mixed with the recombinant GST- MEK as a substrate, and in vitro kinase reactions for B-Raf were performed. Blotting with an anti-B-Raf antibody showed equal protein loading (middle panel). Whole cell lysates (WCL) were blotted with anti-H-Ras antibody to monitor the expression of RasN17 (lower panel). B, the intensity of the GST-MEK phosphorylation by B-Raf immuno- precipitated from mock-transfected (black bar) or RasN17-transfected cells (white bar) was quantified by densitometric analysis. The relative B-Raf activity at 0 min in mock-transfected cells was assigned to be 1.0. Essentially similar results were obtained in three independent experi- ments. IP, immunoprecipitation.

Article Snippet: Anti-human IL-2 antibodies were from R & D Systems (Minneapolis, MN).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Expressing, Recombinant, In Vitro, Phospho-proteomics, Activity Assay, Immunoprecipitation

Journal: Scientific Reports

Article Title: Cholangiocarcinoma derived exosomes attenuate the anti-tumor functions of NK cells

doi: 10.1038/s41598-026-36706-9

Figure Lengend Snippet: Effect of RBE-Exos on the adhesion of NK-92 cells. ( A ) Growth status of NK-92 cells. NK cells treated with RBE-Exos lost the normal cluster growing. ( B ) CD11a protein levels. ( C ) CD18 protein levels. ( D ) CD54 protein levels. Western blot showed that the protein levels of CD11a, CD18 and, CD54 were obviously decreased in NK-92 cells treated with RBE-Exos. ( E ) Relative transcriptional levels of CD11a , CD18 , CD54 and, CD2 in NK-92 cells. qRT-PCR displayed that RBE-Exos downregulated the transcriptional levels of CD11a , CD18 , CD54 , but not CD2 . Data were presented as mean ± SEM for three independent experiments. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: The primary antibodies were as follows: rabbit anti-human CD63, BCL-2, BCL-XL, CD11a, β-actin (Abcam, USA), mouse anti-human HSP70 (Santa-Cruz, USA), rabbit anti-human CD18 (Proteintech, China), mouse anti-human CD54 (Proteintech, China).

Techniques: Western Blot, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Cholangiocarcinoma derived exosomes attenuate the anti-tumor functions of NK cells

doi: 10.1038/s41598-026-36706-9

Figure Lengend Snippet: Cytotoxicity and expression of adhesion molecules in primary NK cells. ( A ) Cytotoxicity of primary NK cells from CCA patients and healthy people against target cells. LDH detection displayed the cytotoxicity of NK cells from CCA patients was significantly lower than that of NK cells healthy people. Data are presented as the mean ± SEM from n = 3 independent experiments. ( B ) Protein levels of CD18 in primary NK cells from CCA patients and healthy people. Data are presented as the mean ± SEM from n = 3 independent experiments. ( C ) Protein levels of CD54 in primary NK cells from CCA patients and healthy people. Data are presented as the mean ± SEM from n = 3 independent experiments. ** P < 0.01.

Article Snippet: The primary antibodies were as follows: rabbit anti-human CD63, BCL-2, BCL-XL, CD11a, β-actin (Abcam, USA), mouse anti-human HSP70 (Santa-Cruz, USA), rabbit anti-human CD18 (Proteintech, China), mouse anti-human CD54 (Proteintech, China).

Techniques: Expressing

Journal:

Article Title: Cellular Sources of Transforming Growth Factor-? Isoforms in Early and Chronic Radiation Enteropathy

doi:

Figure Lengend Snippet: Intestinal wall immunoreactivity for TGF-β1 (anti-CC), TGF-β2, and TGF-β3 2 weeks (left) and 26 weeks (right) after 21-Gy single-dose irradiation. Very prominent TGF-β1 immunoreactivity and minimal TGF-β2 and TGF-β3 immunoreactivity is seen at both observation times.

Article Snippet: 31 Antibodies to pan-TGF-β (AB-100-NA), TGF-β2 (AB-12-NA), and TGF-β3 (AB-244-NA) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Irradiation

Journal:

Article Title: Cellular Sources of Transforming Growth Factor-? Isoforms in Early and Chronic Radiation Enteropathy

doi:

Figure Lengend Snippet: In situ hybridization demonstrating TGF-β1 mRNA expression in irradiated intestine. A: TGF-β1 expression in regenerating crypt at the edge of a radiation-induced mucosal ulcer 2 weeks after 21-Gy single-dose irradiation. Magnification, ×200; bar, 100 μm. B: Inflammatory cells and fibroblast-like cells in the base of a radiation-induced mucosal ulcer expressing TGF-β1 2 weeks after 21-Gy single-dose irradiation. Magnification, ×400; bar, 50 μm. C: Intestinal wall beneath a radiation-induced mucosal ulcer (right overview; magnification, ×100; bar, 100 μm). Strong TGF-β1 expression in smooth muscle cells (left upper panel; magnification, ×400) and peritoneal mesothelium (left lower panel; magnification, ×400) is seen 2 weeks after 21-Gy single-dose irradiation. D: TGF-β1 expression in vascular endothelial cells and perivascular cells 26 weeks after 21-Gy single-dose irradiation. Magnification, ×400; bar, 50 μm. E: Fibroblasts in fibrotic area expressing TGF-β1 mRNA 26 weeks after 21-Gy single-dose irradiation. Magnification, ×400; bar, 50 μm.

Article Snippet: 31 Antibodies to pan-TGF-β (AB-100-NA), TGF-β2 (AB-12-NA), and TGF-β3 (AB-244-NA) were purchased from R&D Systems (Minneapolis, MN).

Techniques: In Situ Hybridization, Expressing, Irradiation

Journal:

Article Title: Cellular Sources of Transforming Growth Factor-? Isoforms in Early and Chronic Radiation Enteropathy

doi:

Figure Lengend Snippet: Expression of TGF-β mRNA in Sham-Irradiated and Irradiated Intestine, 2 and 26 Weeks after Irradiation

Article Snippet: 31 Antibodies to pan-TGF-β (AB-100-NA), TGF-β2 (AB-12-NA), and TGF-β3 (AB-244-NA) were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Irradiation