antibodies escc cell lines  (ATCC)

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