Journal: Frontiers in Cell and Developmental Biology

Article Title: CLIC1 and CLIC4 demonstrate cell protective antioxidant activity against UV exposure

doi: 10.3389/fcell.2025.1674374

Figure Lengend Snippet: Western blot and densitometry analysis of CLIC siRNA knockdowns or rCLICs treated HDF whole cell lysates either non-treated (No UV) or exposed to 30 min of UV irradiation (+UV). [i] Western blot sections showing different antibody staining from one representative gel. Densitometry analysis for each of the different antibodies as shown in the subsequent figures [ii] CLIC1; [iii] CLIC4; [iv] Nitrotyrosine; [v] Catalse; [vi] SOD1 and [vii] Thioredoxin. In each figure, the results are shown for recombinant CLIC treated HDF cells: rCLIC1 (pink) and rCLIC4 (green); siRNA knockdown cells: CLIC1-KD (red), CLIC4-KD (blue) and CLIC1 and CLIC4 Double Knockdown (DKD, orange). The controls either non-treated HDF cells are shown in grey and the Scramble Control (Scmb C), treated with equimolar scramble siRNA, is shown in silver. 10 μg of protein was loaded for each sample. Data expressed as Mean ± SEM. Two-way ANOVA with Tukey’s multiple comparisons test was done. *P < 0.05, Cell lysates was collected from three different passages with WB analysis done on each passage. The original full-length gels (and the replicates) are shown in , where the gel sections provided in this image are highlighted in a red box.

Article Snippet: In order to detect changes in the expression levels of different cellular oxidative stress markers, the following antibodies were used: Catalase (sc-271803), Thioredoxin (Trx, sc-271281), Nitrotyrosine (sc-32757), Superoxide Dismutase type 1 (SOD1, sc-101523) (all antibodies were from Santa Cruz and used in a 1:1000 dilution in PBS-T), with β-Actin (Invitrogen) used as the loading control at a 1:2500 dilution.

Techniques: Western Blot, Irradiation, Staining, Recombinant, Knockdown, Control

Journal: Gastroenterology Report

Article Title: Targeting transglutaminase 2: pathways to celiac disease therapies

doi: 10.1093/gastro/goaf086

Figure Lengend Snippet: SiRNA knockdown of ERp57, TRX, and TG2, and the effect on cellular/extracellular TG2 activity. Caco-2 cells were incubated with specific siRNAs or mock siRNA for 48 h. The efficacy of the siRNA knockdown was determined by using Western blotting. Expression of ERp57, TRX, and TG2 was normalized against the housekeeping protein GAPDH. (A–C) SiRNA treatment significantly reduced expression of (A) ERp57, (B) TRX, and (C) TG2. Incubation with mock siRNA did not affect protein expression ( n = 4). (D) Treatment with ERp57-siRNA und TRX-siRNA did not show any off-target effects on TG2 expression ( n = 3). (E) Cellular expression of ERp57, TRX, and TG2 determined by fluorometry after siRNA knockdown. Protein levels of all three target proteins were reduced by siRNA-mediated knockdown. (F) Cellular TG2 activity was determined by fluorometry after siRNA treatment. SiRNA knockdown of TRX and TG2 reduced cellular TG2 activity ( n = 5). (G) Extracellular expression of ERp57, TRX, and TG2 was investigated in unpermeabilized Caco-2 cells by fluorometry. Protein levels of all three target proteins were significantly reduced by siRNA-mediated knockdown. (H) Extracellular TG2 activity was investigated in unpermeabilized Caco-2 cells by fluorometry. SiRNA knockdown of TRX and TG2 significantly reduced extracellular TG2 activity ( n = 5). All data are shown as mean ± SD. Statistical significance was tested by using Student’s t -test with Welch correction where appropriate. * P < 0.05, ** P < 0.01, *** P < 0.001. GAPDH = glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: In brief, lipofectamine 2000 (1:100; 11668019; Thermo Fisher Scientific) and the corresponding siRNAs (1:50; mock siRNA, ERp57 -siRNA, TRX -siRNA, and TG2 -siRNA from Santa Cruz Biotechnology, Dallas, TX, USA) were diluted in Opti-MEM medium (Thermo Fisher Scientific).

Techniques: Knockdown, Activity Assay, Incubation, Western Blot, Expressing

Journal: Egyptian Journal of Basic and Applied Sciences

Article Title: Thioredoxin-1 and MMP-9 as biomarkers in breast cancer metastasis in Egyptian female patients

doi: 10.1016/j.ejbas.2018.01.001

Figure Lengend Snippet: Fig. 4. Positive correlation between serum thioredoxin-1 (Trx-1) level and matrix metalloproteinase-9 (MMP-9) level in all breast cancer patients.

Article Snippet: Serum Trx-1 level and MMP-9 level were measured by commercially available enzyme-linked immunosorbent assay (ELISA) kits using human thioredoxin ELISA kit, (Boster bio, Pleasanton, CA, USA) and human MMP-9 platinum ELISA kit, (eBioscience, San Diego, CA, USA).

Techniques:

Journal: Egyptian Journal of Basic and Applied Sciences

Article Title: Thioredoxin-1 and MMP-9 as biomarkers in breast cancer metastasis in Egyptian female patients

doi: 10.1016/j.ejbas.2018.01.001

Figure Lengend Snippet: Fig. 5. Positive correlation between serum thioredoxin-1 (Trx-1) and cancer antigen 15-3 (CA15-3) levels (a) and serum Trx-1 and carcinoembryonic antigen (CEA) levels (b) in all breast cancer patients.

Article Snippet: Serum Trx-1 level and MMP-9 level were measured by commercially available enzyme-linked immunosorbent assay (ELISA) kits using human thioredoxin ELISA kit, (Boster bio, Pleasanton, CA, USA) and human MMP-9 platinum ELISA kit, (eBioscience, San Diego, CA, USA).

Techniques:

Journal: Egyptian Journal of Basic and Applied Sciences

Article Title: Thioredoxin-1 and MMP-9 as biomarkers in breast cancer metastasis in Egyptian female patients

doi: 10.1016/j.ejbas.2018.01.001

Figure Lengend Snippet: Fig. 7. Receiver operating characteristic curve (ROC) for serum thioredoxin-1 (Trx-1) level (a) and for serum matrix metalloproteinase-9 (MMP-9) level (b) for detection of breast cancer metastases.

Article Snippet: Serum Trx-1 level and MMP-9 level were measured by commercially available enzyme-linked immunosorbent assay (ELISA) kits using human thioredoxin ELISA kit, (Boster bio, Pleasanton, CA, USA) and human MMP-9 platinum ELISA kit, (eBioscience, San Diego, CA, USA).

Techniques: