te4  (ATCC)

Journal: bioRxiv

Article Title: CD8 + T cells are primed by cDC1 and exacerbate tau-mediated neurodegeneration

doi: 10.64898/2026.02.26.708260

Figure Lengend Snippet: ( a ) Representative images of 9.5-month-old male (upper) and female (lower) E4 WT , E4 Δ+32 , TE4 WT , and TE4 Δ+32 mice. ( b ) Volumetric analysis of the hippocampus, piriform entorhinal cortex and lateral ventricles of male (upper) and female (lower) E4 WT (n=7 for male and n=8 for female), E4 Δ+32 (n=9 for both sexes), TE4 WT (n=19 for male and n=22 for female), and TE4 Δ+32 (n=21 for both sexes) mice. ( c ) Representative images showing the thickness of the dentate gyrus (DG) granule cell layer in four strains of mice. ( d ) DG thickness analysis in four strains of mice. ( e ) Level of plasma neurofilament light chain (NfL) in male or female TE4 WT and TE4 Δ+32 mice. Data are presented as mean values ± SEMs. Statistical significance was defined using two-way ANOVA with Šídák’s multiple comparisons test ( b and d ) and Mann-Whitney test, two-tailed ( e ). P value (actual value) was indicated in the figure. ****P<0.0001; NS, no significance.

Article Snippet: The TE4 mice were then crossed with Irf8 +32 -/- mice with B57BL/6 background (Stock No. 032744, Jackson Laboratories) for two generations to generate E4 WT , E4 Δ+32 , TE4 WT , TE4 Δ+32 mice.

Techniques: Clinical Proteomics, MANN-WHITNEY, Two Tailed Test

Journal: bioRxiv

Article Title: CD8 + T cells are primed by cDC1 and exacerbate tau-mediated neurodegeneration

doi: 10.64898/2026.02.26.708260

Figure Lengend Snippet: ( a ) Representative immunofluorescent images of reactive microglia from four strains of the male mice at 9.5-month-old. The hippocampal sections were stained with IBA1 (Red), MHC-II (Green), and P2RY12 (Magenta). ( b ) Percent of the area analysis covered by IBA1 (Left), MHC-II (Middle), and P2RY12 (Right). ( c ) Representative immunofluorescent images of reactive astrocytes from four strains of the male mice at 9.5-month-old. The hippocampal sections were stained with Vimentin (Red), GFAP (Green), and DAPI (Blue). ( d ) Percent area covered by GFAP (Left) and Vimentin (Middle) staining. ( e ) Representative images showing the co-localization of Vimentin with GFAP from hippocampal sections from TE4 WT or TE4 Δ+32 mice. ( f ) Percentage of Vimentin staining overlapping with GFAP staining from the sections from the indicated mice. n=7 for E4 WT , n=9 for E4 Δ+32 , n=19 for TE4 WT , and n=21 for TE4 Δ+32 mice. Data are presented as mean values ± SEMs. Statistical significance was defined using two-way ANOVA with Šídák’s multiple comparisons test. P value (actual value) was indicated in the figure. Scale bar, 25μm. ****P<0.0001; NS, no significance.

Article Snippet: The TE4 mice were then crossed with Irf8 +32 -/- mice with B57BL/6 background (Stock No. 032744, Jackson Laboratories) for two generations to generate E4 WT , E4 Δ+32 , TE4 WT , TE4 Δ+32 mice.

Techniques: Staining

Journal: bioRxiv

Article Title: CD8 + T cells are primed by cDC1 and exacerbate tau-mediated neurodegeneration

doi: 10.64898/2026.02.26.708260

Figure Lengend Snippet: ( a ) UMAP plot shows individual clusters in CD45 hi leukocytes from brains of male E4 WT , E4 Δ+32 , TE4 WT , and TE4 Δ+32 mice. ( b ) Dot plot identifies 11 clusters of leukocytes. ( c ) Representative flow cytometry plot confirms the identity of different leukocyte populations. ( d ) UMAP (left) plot reveals the population shift comparing the four strains of mice. Bar graph (right) shows the frequency of cells populating individual clusters. ( e ) Representative flow cytometry analysis identifying lymphocytes including conventional T cells (CD4 T cells and CD8 T cells), NK cells, and NKT cells in hippocampus and cortex of brains from 9.5-month-old male E4 WT , E4 Δ+32 , TE4 WT , and TE4 Δ+32 mice. ( f-k ) Frequency of CD45 hi ( f ), αβ T cells ( g ), CD4 T cells ( h ), CD8 T cells ( i ), NK cells ( j ), and NKT cells ( k ) from hippocampus and cortex from 9.5-month-old male E4 WT (n=14), E4 Δ+32 (n=11), TE4 WT (n=17), and TE4 Δ+32 (n=15) mice. ( l ) Representative immunofluorescence images of T cell accumulation in hippocampal regions of brains from the four strains of male mice. The hippocampal sections were stained with CD3 (Green), CD4 (Red), CD8 (Magenta), and IBA1 (Blue). Scale bar: 40μm. ( m-p ) Data summarizes the number of the CD3+ total T cells ( m ), CD8+ T cells ( n ), CD4+ T cells ( o ), and CD3+CD4-CD8- T cells ( p ) within the hippocampus. Male E4 WT (n=7), male E4 Δ+32 (n=9), male TE4 WT (n=19), and male TE4 Δ+32 (n=21). Data are presented as mean values ± SEMs. Statistical significance was defined using two-way ANOVA with Šídák’s multiple comparisons test ( f-k ) and unpaired student t test, two tailed ( m-p ). P value (actual value) was indicated in the figure. ****P<0.0001; NS, no significance.

Article Snippet: The TE4 mice were then crossed with Irf8 +32 -/- mice with B57BL/6 background (Stock No. 032744, Jackson Laboratories) for two generations to generate E4 WT , E4 Δ+32 , TE4 WT , TE4 Δ+32 mice.

Techniques: Flow Cytometry, Immunofluorescence, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: CD8 + T cells are primed by cDC1 and exacerbate tau-mediated neurodegeneration

doi: 10.64898/2026.02.26.708260

Figure Lengend Snippet: ( a ) UMAP plot shows individual clusters in enriched CD3 + T cells from brains of E4 WT , E4 Δ+32 , TE4 WT , and TE4 Δ+32 mice. ( b ) Feature plots show the expression of specific markers in clusters of cells. ( c ) Density plot (upper) shows the changes of the frequencies for each cluster and bar graph calculates the proportion of each cluster in CD4 + T cells (lower left) and CD8 + T cells (lower right). ( d ) Representative flow cytometry identifies naïve T cells (upper left) and Tregs (lower left) and frequencies of each population in CD4 + T cells among the four strains of male mice (right). ( e ) Representative flow cytometry identifies naïve T cells (upper left) and exhausted age associated T cells (lower left) and frequencies of each population in CD8 + T cells among the four strains of male mice (right). ( f ) UMAP plot shows the top T cell clonotypes in each strain of mice. Red dots represent top clones and size of the dots represented the proportion of the clonotypes among all T cells with paired TCR information (left). ( g ) Gini index bar plots of clonotypes in different cell types under different conditions. Higher Gini index indicates less even frequency of clonotypes and greater extent of clonal expansion. Data are presented as mean values ± SEMs. Statistical significance was defined using two-way ANOVA with Šídák’s multiple comparisons test ( d and e ). P value (actual value) was indicated in the figure. ****P<0.0001; NS, no significance.

Article Snippet: The TE4 mice were then crossed with Irf8 +32 -/- mice with B57BL/6 background (Stock No. 032744, Jackson Laboratories) for two generations to generate E4 WT , E4 Δ+32 , TE4 WT , TE4 Δ+32 mice.

Techniques: Expressing, Flow Cytometry, Clone Assay

Journal: Clinics (Sao Paulo, Brazil)

Article Title: The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

doi: 10.1016/j.clinsp.2023.100276

Figure Lengend Snippet: Fig. 3. TMEM26 depletion in ESCC cells suppressed EMT-related alterations. (A) Transfection of siRNA targeting the TMEM26 gene (three independent targets) effi- ciently downregulated TMEM26 expression in TMEM26-high KYSE270 and TE8 cells, as detected in western blotting. (B) Transwell assay revealed a reduced number of invaded cells with TMEM26 RNAi, compared with control RNAi, in TMEM26-high KYSE270, and TE8 cells. Left: representative images. Right: quantification and sta- tistical result. Scale bar = 100 µm. (C) Wound healing assay for TMEM26 RNAi KYSE270 and TE8 cells showed fewer cells migrating into the wound region, compared with control RNAi cells. Left: representative images. Right: quantification and statistical result. Scale bar = 100 µm. (D) Western blotting to detect mesenchymal and epithelial marker expressions showed downregulated twist, snail, N-cadherin, and vimentin but upregulated E-cadherin in TMEM26 RNAi KYSE270 and TE8 cells. Data are presented as means ± S.D., n = 3, ***p < 0.001.

Article Snippet: Cell lines, reagents, and antibodies ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA).

Techniques: Transfection, Expressing, Western Blot, Transwell Assay, Control, Wound Healing Assay, Marker

Journal: Clinics (Sao Paulo, Brazil)

Article Title: The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

doi: 10.1016/j.clinsp.2023.100276

Figure Lengend Snippet: Fig. 2. TMEM26 expression and cellular localization in various ESCC cell lines. (A) The abundance of TMEM26 in the normal esophageal epithelial cell line (HET-1A) and a collection of ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) were examined by western blotting. (B) Immunofluorescent staining of TMEM26 with DAP in KYSE270 and TE8 cells verified the plasma membrane localization of TMEM26. Scale bar = 10 µm.

Article Snippet: Cell lines, reagents, and antibodies ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA).

Techniques: Expressing, Western Blot, Staining, Clinical Proteomics, Membrane

Journal: Clinics (Sao Paulo, Brazil)

Article Title: The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

doi: 10.1016/j.clinsp.2023.100276

Figure Lengend Snippet: Fig. 4. TMEM26 overexpression in ESCC cells promoted EMT-related alterations. (A) Transfection of TMEM26-expressing plasmid by lentiviral vectors efficiently over- regulated TMEM26 in TMEM26-low KYSE150 and KYSE450 cells, as shown by western blotting. (B) Transwell assay showed a higher number of invaded cells with TMEM26 overexpression than with control transfection in TMEM26-low KYSE150 and KYSE450 cells. Left: representative images. Right: quantification and statistical result. Scale bar = 100 µm. (C) The wound healing assay showed that the number of cells migrating into the wound region in TMEM26-overexpressing KYSE150 and KYSE450 cells was higher than that in control transfection cells. Left: representative images. Right: quantification and statistical result. (D) Western blotting detection of mesenchymal and epithelial marker expressions showed upregulated twist, snail, N-cadherin, and vimentin but downregulated E-cadherin in TMEM26-overexpress- ing KYSE150 and KYSE450 cells than that in control transfection cells. Data are presented as means ± S.D., n = 3, ***p < 0.001.

Article Snippet: Cell lines, reagents, and antibodies ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA).

Techniques: Over Expression, Transfection, Expressing, Plasmid Preparation, Western Blot, Transwell Assay, Control, Wound Healing Assay, Marker

Journal: Clinics (Sao Paulo, Brazil)

Article Title: The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

doi: 10.1016/j.clinsp.2023.100276

Figure Lengend Snippet: Fig. 5. TMEM26 did not affect ESCC cell growth. (A) In TE8 (left) and KYSE270 (right) cells, siRNA knockdown of TMEM26 (three independent targets) did not show observable differences in the growth curve compared with siRNA control cells, as determined by MTT assay. (B) In KYSE150 (left) and KYSE450 (right) cells, TMEM26 overexpression due to lentiviral transfection did not show observable differences in the growth curve compared with control cells, as determined by MTT assay. Data are presented as means ± S.D., n = 6.

Article Snippet: Cell lines, reagents, and antibodies ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA).

Techniques: Knockdown, Control, MTT Assay, Over Expression, Transfection

Journal: Clinics (Sao Paulo, Brazil)

Article Title: The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

doi: 10.1016/j.clinsp.2023.100276

Figure Lengend Snippet: Fig. 7. Detection for NF-κB signaling activation by TMEM26 and the determining effects of NF-κB signaling on EMT in ESCC cells. (A) In TMEM26-high TE8 (left) and KYSE270 (right) cells, siRNA knockdown of TMEM26 (three independent targets) did not show observable differences for the total number of p65 and IκBα; however, the p65 and IκBα phosphorylation was dra- matically declined, as detected in western blotting. (B) TMEM26 overexpression in TMEM26-low KYSE150 and KYSE450 cells resulted in elevated p65 and IκBα phosphorylation, whereas the total amount of p65 and IκBα remained comparable, as detected in western blotting. (C) In KYSE150 and KYSE450 cells, TMEM26 overexpression enhanced the expression of twist; how- ever, the treatment with PS1145, an NF-κB inhibitor, blocked this effect, as detected in western blotting. (D) Wound healing assay for TMEM26-overexpressing KYSE150 and KYSE450 cells showed a higher number of cells migrating into the wound region, which was blocked by PS1145. Left: representative images. Right: quantification and statistical result. Scale bar = 100 µm. (E) Transwell assay for KYSE150 and KYSE450 cells showed a higher number of invading cells after overexpressing TMEM26, which was blocked by PS1145. Right: quantification and statisti- cal result. Scale bar = 100 µm. Data are presented as means ± S.D., n = 3, *p < 0.05, **p < 0.01, and ***p < 0.001 versus the vector group, ###p < 0.001 vs. the TMEM26 OE group.

Article Snippet: Cell lines, reagents, and antibodies ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA).

Techniques: Activation Assay, Knockdown, Phospho-proteomics, Western Blot, Over Expression, Expressing, Wound Healing Assay, Transwell Assay, Plasmid Preparation

Journal: Clinics (Sao Paulo, Brazil)

Article Title: The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

doi: 10.1016/j.clinsp.2023.100276

Figure Lengend Snippet: Fig. 8. Regulation of plasma membrane distribution of TJ proteins by TMEM26 in TE8 and KYSE150 ESCC cells. (A and B) Immunofluorescent staining for claudin-1o Occludin, and ZO-1 with DAPI in TE8 cells showed their plasma membrane localization. Compared with the siRNA control, the TE8 cells with siRNA knockdown of TMEM26 (three independent targets) elevated the distribution of claudin-1, occludin, and ZO-1 at the plasma membrane. Quantification of plasma membrane signal intensity is shown in (B). (C and D) Immunofluorescent staining for claudin-1, occludin, and ZO-1 with DAPI in KYSE150 cells showed their plasma membrane localiza- tion. Compared with control transfection, KYSE150 cells with overexpressed TMEM26 showed a reduced distribution of claudin-1, occludin, and ZO-1 at the plasma membrane. Quantification of plasma membrane signal intensity is shown in (D). Scale bar = 10 µm. Data are presented as means ± S.D., n = 3, **p < 0.01, and ***p < 0.001.

Article Snippet: Cell lines, reagents, and antibodies ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA).

Techniques: Clinical Proteomics, Membrane, Staining, Control, Knockdown, Transfection