Review




Structured Review

Proteintech zo 1
Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B <t>(ZO‐1</t> signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.
Zo 1, supplied by Proteintech, used in various techniques. Bioz Stars score: 97/100, based on 2559 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+zo-1/ZO-1+Antibody/pmc13650023-29-30-32
Average 97 stars, based on 2559 article reviews
zo 1 - by Bioz Stars, 2026-10
97/100 stars

Images

1) Product Images from "Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD"

Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD

Journal: Advanced Science

doi: 10.1002/advs.78081

Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B (ZO‐1 signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.
Figure Legend Snippet: Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B (ZO‐1 signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.

Techniques Used: Cell Culture, Immunofluorescence, Staining, Expressing, EdU Assay, Comparison, MANN-WHITNEY, Control

Effects of MMC and CDDP on Functional Marker Protein Expression in Primary Cultured Rat PAECs, PVECs, and PMVECs. (A–L) Immunoblots and analyzed graphs showing the protein expression of ZO‐1, VEGFR2, FANCL, PCNA, Slug, Bcl2, Bax, and γ‐H2AX in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Data are presented as mean ± SD, n = 5 rats per group. Data are presented as mean ± SD. Differences between two groups were analyzed by unpaired t ‐test, except for D (ZO1 at 48 h and VEGFR2 at 24 and 48 h), F (Bax/Bcl2 and γ‐H2AX at 24 h and Slug at 48 h), H (ZO1 at 24 h and PCNA at 48 h), and J (γ‐H2AX at 24 h), for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. (M, N) Heatmap of RNA‑seq data depicting gene‑ and pathway‑level changes in cultured rat PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Veh: vehicle control.
Figure Legend Snippet: Effects of MMC and CDDP on Functional Marker Protein Expression in Primary Cultured Rat PAECs, PVECs, and PMVECs. (A–L) Immunoblots and analyzed graphs showing the protein expression of ZO‐1, VEGFR2, FANCL, PCNA, Slug, Bcl2, Bax, and γ‐H2AX in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Data are presented as mean ± SD, n = 5 rats per group. Data are presented as mean ± SD. Differences between two groups were analyzed by unpaired t ‐test, except for D (ZO1 at 48 h and VEGFR2 at 24 and 48 h), F (Bax/Bcl2 and γ‐H2AX at 24 h and Slug at 48 h), H (ZO1 at 24 h and PCNA at 48 h), and J (γ‐H2AX at 24 h), for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. (M, N) Heatmap of RNA‑seq data depicting gene‑ and pathway‑level changes in cultured rat PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Veh: vehicle control.

Techniques Used: Functional Assay, Marker, Expressing, Cell Culture, Western Blot, MANN-WHITNEY, Control

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Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B <t>(ZO‐1</t> signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.
Anti Zo 1, supplied by Proteintech, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Boster Bio anti zo 1
Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B <t>(ZO‐1</t> signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.
Anti Zo 1, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+zo-1/Anti-ZO1+tight+junction+protein%2FTJP1+Antibody+Picoband/pm42805596-126-52-56
Average 94 stars, based on 1 article reviews
anti zo 1 - by Bioz Stars, 2026-10
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Proteintech antibodies against zo 1
Primer sequences for target genes and the reference gene.
Antibodies Against Zo 1, supplied by Proteintech, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+zo-1/ZO-1+Antibody/pmc13649658-70-30-34
Average 97 stars, based on 1 article reviews
antibodies against zo 1 - by Bioz Stars, 2026-10
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Image Search Results


ABCA1 deficiency disrupts the junctional integrity of the colonic mucosal barrier. (A) Representative images of immunofluorescence staining of the zippering junction with E‐cadherin. (B, C) Effects of ABCA1 silencing on cholesterol crystals (CHO)‐induced cell injury. Caco‐2 cells were transfected with siNC or siABCA1 for 24 h, and then exposed to 1 mmol/L CHO or not for 24 h. The cell viability was determined using Calcein‐AM/PI staining (B) and CCK8 assay (C). The data in panel C are expressed as mean ± SD ( n = 4; *** p < 0.001 vs . siNC without CHO by Student's t ‐test). (D) ABCA1 silencing aggravates CHO‐mediated barrier dysfunction in Caco‐2 cells. Caco‐2 cells were transfected with siNC or siABCA1 for 24 h, and then exposed to 1 mmol/L CHO for 6 h. Cellular lysates were assayed for E‐cadherin, Occludin and ZO‐1 expression. The respective densitometric quantitations of bands are presented in the bar graphs to the right of the blots. Data are expressed as mean ± SD ( n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 vs . siNC without CHO by Student's t ‐test).

Journal: Animal Models and Experimental Medicine

Article Title: ABCA1 deficiency‐induced cholesterol accumulation aggravates colonic epithelial injury in knockout hamsters

doi: 10.1002/ame2.70301

Figure Lengend Snippet: ABCA1 deficiency disrupts the junctional integrity of the colonic mucosal barrier. (A) Representative images of immunofluorescence staining of the zippering junction with E‐cadherin. (B, C) Effects of ABCA1 silencing on cholesterol crystals (CHO)‐induced cell injury. Caco‐2 cells were transfected with siNC or siABCA1 for 24 h, and then exposed to 1 mmol/L CHO or not for 24 h. The cell viability was determined using Calcein‐AM/PI staining (B) and CCK8 assay (C). The data in panel C are expressed as mean ± SD ( n = 4; *** p < 0.001 vs . siNC without CHO by Student's t ‐test). (D) ABCA1 silencing aggravates CHO‐mediated barrier dysfunction in Caco‐2 cells. Caco‐2 cells were transfected with siNC or siABCA1 for 24 h, and then exposed to 1 mmol/L CHO for 6 h. Cellular lysates were assayed for E‐cadherin, Occludin and ZO‐1 expression. The respective densitometric quantitations of bands are presented in the bar graphs to the right of the blots. Data are expressed as mean ± SD ( n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001 vs . siNC without CHO by Student's t ‐test).

Article Snippet: Then, proteins were transferred to a polyvinylidene difluoride membrane and hybridized with the following antibodies: ABCA1 (26564‐1‐AP, Proteintech, Wuhan, China), ABCA1 (96292, Cell Signaling Technology, Danvers, MA), ZO‐1 (A0659, ABclonal, Wuhan, China), E‐cadherin (sc‐8426, Santa cruz Biotechnology, Santa Cruz, CA), Occludin (66378‐1‐Ig, Proteintech, Wuhan, China), p‐p38 (8796‐1‐AP, Proteintech, Wuhan, China) or β‐actin (66009‐1‐Ig, Proteintech, Wuhan, China).

Techniques: Immunofluorescence, Staining, Transfection, CCK-8 Assay, Expressing

Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B (ZO‐1 signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.

Journal: Advanced Science

Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD

doi: 10.1002/advs.78081

Figure Lengend Snippet: Effects of MMC and CDDP on Barrier Junction, Proliferation, and Tube Formation in Primarily Cultured Rat PAECs, PVECs, and PMVECs. (A, B) Immunofluorescence staining and quantification of ZO‑1 expression in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 7 independent experiments for PAECs and n = 10 for PVECs and PMVECs. (C, D) Edu incorporation and quantification in PAECs, PVECs and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 5 independent experiments per group. (E, F) Tube formation images and quantification of the angiogenic capacity in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. n = 6 independent experiments per group. Data are presented as mean ± SD. Differences for B (ZO‐1 signal intensity) and D (EdU‐positive cell percentage) were analyzed by one‐way ANOVA with Tukey's HSD post hoc test. Differences between two groups in F were analyzed by unpaired t ‐test, except for the comparison of total segment length between Veh‐ and MMC‐treated PMVECs, for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. Scale bar = 50 µm in A and C, and 100 µm in E. Veh: vehicle control.

Article Snippet: Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining.

Techniques: Cell Culture, Immunofluorescence, Staining, Expressing, EdU Assay, Comparison, MANN-WHITNEY, Control

Effects of MMC and CDDP on Functional Marker Protein Expression in Primary Cultured Rat PAECs, PVECs, and PMVECs. (A–L) Immunoblots and analyzed graphs showing the protein expression of ZO‐1, VEGFR2, FANCL, PCNA, Slug, Bcl2, Bax, and γ‐H2AX in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Data are presented as mean ± SD, n = 5 rats per group. Data are presented as mean ± SD. Differences between two groups were analyzed by unpaired t ‐test, except for D (ZO1 at 48 h and VEGFR2 at 24 and 48 h), F (Bax/Bcl2 and γ‐H2AX at 24 h and Slug at 48 h), H (ZO1 at 24 h and PCNA at 48 h), and J (γ‐H2AX at 24 h), for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. (M, N) Heatmap of RNA‑seq data depicting gene‑ and pathway‑level changes in cultured rat PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Veh: vehicle control.

Journal: Advanced Science

Article Title: Alkylating Agent–Induced FANCL Impairment Damages Pulmonary Arterial, Venous, and Microvascular Endothelial Cells and Predisposes to PAH/PVOD

doi: 10.1002/advs.78081

Figure Lengend Snippet: Effects of MMC and CDDP on Functional Marker Protein Expression in Primary Cultured Rat PAECs, PVECs, and PMVECs. (A–L) Immunoblots and analyzed graphs showing the protein expression of ZO‐1, VEGFR2, FANCL, PCNA, Slug, Bcl2, Bax, and γ‐H2AX in PAECs, PVECs, and PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Data are presented as mean ± SD, n = 5 rats per group. Data are presented as mean ± SD. Differences between two groups were analyzed by unpaired t ‐test, except for D (ZO1 at 48 h and VEGFR2 at 24 and 48 h), F (Bax/Bcl2 and γ‐H2AX at 24 h and Slug at 48 h), H (ZO1 at 24 h and PCNA at 48 h), and J (γ‐H2AX at 24 h), for which the Mann–Whitney U‐test was used. A value of P < 0.05 is considered statistically significant, n.s. indicates no statistical significance. (M, N) Heatmap of RNA‑seq data depicting gene‑ and pathway‑level changes in cultured rat PMVECs treated with MMC (0.5 µg/mL) or CDDP (12 µM) for 24 or 48 h. Veh: vehicle control.

Article Snippet: Primary antibodies for fibronectin (FN, #15613‐I‐AP, Proteintech, Rosemont, IL, USA), Twist (ab50887, Abcam, Cambridge, MA, USA), Vimentin (#5741, Cell Signaling Technology, Danvers, MA, USA), Snail (#ab53519, Abcam, Cambridge, MA, USA), ZO‐1 (21773‐1‐AP, proteintech, Wuhan, China), VEGFR2 (9698, CST, Boston, USA), Slug (YM8357, Immunoway, California, USA), proliferating cell nuclear antigen (PCNA, ab29, Abcam, Cambridge, MA, USA), CD31 (AF3628, R&D Systems, Minneapolis, MN, USA), Cleaved caspase3 (#9664, Cell Signaling Technology, Danvers, MA, USA), Bax (#2772S, Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma/leukemia 2 (Bcl‐2, 13–8800, Thermo Fisher Scientific, Rockford, IL, USA), γ‐H2AX (#05‐635, EMD Millipore/Merck, USA), α‐smooth muscle actin (α‐SMA, ab124964 and ab7817, Abcam, Cambridge, MA, USA), FANCL (LS‐C807939, LifeSpan BioSciences, Seattle, WA, USA), FANCD2 (NB100‐182, Novus Biologicals, USA), general control nonderepressible 2 (GCN2, #46041, Cell Signaling Technology, Danvers, MA, USA), and β‐actin (66009‐1‐Ig, Proteintech, Rosemont, IL, USA) were used for immunoblotting, immunofluorescence (IF), and immunohistochemistry (IHC) staining.

Techniques: Functional Assay, Marker, Expressing, Cell Culture, Western Blot, MANN-WHITNEY, Control

Primer sequences for target genes and the reference gene.

Journal: Frontiers in Microbiology

Article Title: Tight junction suppression by candidalysin promotes gut-to-blood dissemination of Candida albicans

doi: 10.3389/fmicb.2026.1919478

Figure Lengend Snippet: Primer sequences for target genes and the reference gene.

Article Snippet: The membranes were blocked with 5% bovine serum albumin in Tris-buffered saline containing 0.1% Tween-20 for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies against ZO-1 (rabbit, Proteintech, Cat. No. 21773-1-AP, 1:1000), occludin (rabbit, Proteintech, Cat. No. 27260-1-AP, 1:1000), claudin-1 (rabbit, Proteintech, Cat. No. 28674-1-AP, 1:1000), Dsg2 (rabbit, Proteintech, Cat. No. [21880-1-AP], 1:1000), β-actin (rabbit, Proteintech, Cat. No. 20536-1-AP, 1:1000 in NCM-460) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (rabbit, Proteintech, Cat. No. 10494-1-AP, 1:1000 in mice tissues).

Techniques:

Candidalysin disrupts intestinal tight junction proteins. (A) Western blot analysis of intestinal tissues at 48 h post-infection showing expression levels of tight junction proteins ZO-1, Occludin, and Claudin-1. Protein levels were reduced in tissues from mice infected with WT and ECE1-reconstituted strains, whereas expression in ece1Δ/Δ-infected mice was relatively preserved. (B) Immunohistochemical staining of intestinal mucosa for ZO-1, Occludin, and Claudin-1. Loss of protein expression and disrupted mucosal structure were observed in WT (1477) and ECE1-reconstituted (2059) infections, whereas continuous and well-distributed staining was maintained in ece1Δ/Δ (2057)-infected tissues, similar to negative control mice. The staining pattern was consistent with strain-dependent TJ disruption during intestinal infection.

Journal: Frontiers in Microbiology

Article Title: Tight junction suppression by candidalysin promotes gut-to-blood dissemination of Candida albicans

doi: 10.3389/fmicb.2026.1919478

Figure Lengend Snippet: Candidalysin disrupts intestinal tight junction proteins. (A) Western blot analysis of intestinal tissues at 48 h post-infection showing expression levels of tight junction proteins ZO-1, Occludin, and Claudin-1. Protein levels were reduced in tissues from mice infected with WT and ECE1-reconstituted strains, whereas expression in ece1Δ/Δ-infected mice was relatively preserved. (B) Immunohistochemical staining of intestinal mucosa for ZO-1, Occludin, and Claudin-1. Loss of protein expression and disrupted mucosal structure were observed in WT (1477) and ECE1-reconstituted (2059) infections, whereas continuous and well-distributed staining was maintained in ece1Δ/Δ (2057)-infected tissues, similar to negative control mice. The staining pattern was consistent with strain-dependent TJ disruption during intestinal infection.

Article Snippet: The membranes were blocked with 5% bovine serum albumin in Tris-buffered saline containing 0.1% Tween-20 for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies against ZO-1 (rabbit, Proteintech, Cat. No. 21773-1-AP, 1:1000), occludin (rabbit, Proteintech, Cat. No. 27260-1-AP, 1:1000), claudin-1 (rabbit, Proteintech, Cat. No. 28674-1-AP, 1:1000), Dsg2 (rabbit, Proteintech, Cat. No. [21880-1-AP], 1:1000), β-actin (rabbit, Proteintech, Cat. No. 20536-1-AP, 1:1000 in NCM-460) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (rabbit, Proteintech, Cat. No. 10494-1-AP, 1:1000 in mice tissues).

Techniques: Western Blot, Analysis, Infection, Expressing, Immunohistochemical staining, Staining, Negative Control, Disruption

Biphasic modulation of tight junction proteins by candidalysin in NCM-460 colonic epithelial cells. Immunofluorescence analysis of ZO-1, Occludin, and Claudin-1 (200× magnification) following exposure of intestinal epithelial cells to different concentrations of candidalysin. Representative immunofluorescence images are shown for vehicle and candidalysin at the indicated concentrations. At 3.125 μg/mL, junction-associated signals were relatively preserved. At 12.5 and 50 μg/mL, junction-associated signals were reduced and showed altered localization, with Occludin and Claudin-1 particularly affected. Primary antibodies against each individual TJ protein were used at 1:500 dilution, followed by FITC-conjugated goat anti-rabbit secondary antibodies for detection. Nuclei were counterstained with DAPI.

Journal: Frontiers in Microbiology

Article Title: Tight junction suppression by candidalysin promotes gut-to-blood dissemination of Candida albicans

doi: 10.3389/fmicb.2026.1919478

Figure Lengend Snippet: Biphasic modulation of tight junction proteins by candidalysin in NCM-460 colonic epithelial cells. Immunofluorescence analysis of ZO-1, Occludin, and Claudin-1 (200× magnification) following exposure of intestinal epithelial cells to different concentrations of candidalysin. Representative immunofluorescence images are shown for vehicle and candidalysin at the indicated concentrations. At 3.125 μg/mL, junction-associated signals were relatively preserved. At 12.5 and 50 μg/mL, junction-associated signals were reduced and showed altered localization, with Occludin and Claudin-1 particularly affected. Primary antibodies against each individual TJ protein were used at 1:500 dilution, followed by FITC-conjugated goat anti-rabbit secondary antibodies for detection. Nuclei were counterstained with DAPI.

Article Snippet: The membranes were blocked with 5% bovine serum albumin in Tris-buffered saline containing 0.1% Tween-20 for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies against ZO-1 (rabbit, Proteintech, Cat. No. 21773-1-AP, 1:1000), occludin (rabbit, Proteintech, Cat. No. 27260-1-AP, 1:1000), claudin-1 (rabbit, Proteintech, Cat. No. 28674-1-AP, 1:1000), Dsg2 (rabbit, Proteintech, Cat. No. [21880-1-AP], 1:1000), β-actin (rabbit, Proteintech, Cat. No. 20536-1-AP, 1:1000 in NCM-460) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (rabbit, Proteintech, Cat. No. 10494-1-AP, 1:1000 in mice tissues).

Techniques: Immunofluorescence, Analysis

Candidalysin regulates tight junction proteins at both protein and transcript levels in colonic epithelial cells. (A) Representative Western blot analysis of ZO-1, Occludin, Claudin-1, and desmoglein-2 (Dsg2) in NCM-460 cells treated with 3.125 μg/mL or 50 μg/mL candidalysin. GAPDH was used as the loading control, whereas Dsg2 was included as a desmosomal comparison protein. (B–E) Densitometric quantification of TJ proteins from n = 3 independent biological experiments. ZO-1 expression was significantly increased at 3.125 μg/mL but markedly reduced at 50 μg/mL. Claudin-1 showed significant reduction at 50 μg/mL, while Occludin levels remained unchanged at both concentrations. (F) Scanning electron microscopy images showing intact polygonal morphology with preserved junctions in untreated cells versus marked structural collapse of cell structures and junctional disintegration after 50 μg/mL candidalysin exposure. (G) qPCR analysis of ZO-1, Occludin, and Claudin-1 transcripts showing significant downregulation at both low and high candidalysin concentrations, with greatest reduction observed at 50 μg/mL.* p < 0.05; ** p < 0.01.

Journal: Frontiers in Microbiology

Article Title: Tight junction suppression by candidalysin promotes gut-to-blood dissemination of Candida albicans

doi: 10.3389/fmicb.2026.1919478

Figure Lengend Snippet: Candidalysin regulates tight junction proteins at both protein and transcript levels in colonic epithelial cells. (A) Representative Western blot analysis of ZO-1, Occludin, Claudin-1, and desmoglein-2 (Dsg2) in NCM-460 cells treated with 3.125 μg/mL or 50 μg/mL candidalysin. GAPDH was used as the loading control, whereas Dsg2 was included as a desmosomal comparison protein. (B–E) Densitometric quantification of TJ proteins from n = 3 independent biological experiments. ZO-1 expression was significantly increased at 3.125 μg/mL but markedly reduced at 50 μg/mL. Claudin-1 showed significant reduction at 50 μg/mL, while Occludin levels remained unchanged at both concentrations. (F) Scanning electron microscopy images showing intact polygonal morphology with preserved junctions in untreated cells versus marked structural collapse of cell structures and junctional disintegration after 50 μg/mL candidalysin exposure. (G) qPCR analysis of ZO-1, Occludin, and Claudin-1 transcripts showing significant downregulation at both low and high candidalysin concentrations, with greatest reduction observed at 50 μg/mL.* p < 0.05; ** p < 0.01.

Article Snippet: The membranes were blocked with 5% bovine serum albumin in Tris-buffered saline containing 0.1% Tween-20 for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies against ZO-1 (rabbit, Proteintech, Cat. No. 21773-1-AP, 1:1000), occludin (rabbit, Proteintech, Cat. No. 27260-1-AP, 1:1000), claudin-1 (rabbit, Proteintech, Cat. No. 28674-1-AP, 1:1000), Dsg2 (rabbit, Proteintech, Cat. No. [21880-1-AP], 1:1000), β-actin (rabbit, Proteintech, Cat. No. 20536-1-AP, 1:1000 in NCM-460) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (rabbit, Proteintech, Cat. No. 10494-1-AP, 1:1000 in mice tissues).

Techniques: Western Blot, Analysis, Control, Comparison, Expressing, Scanning Electron Microscopy, Morphology