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
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![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] <t>Thioredoxin.</t> 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.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_9148/pmc12669148/pmc12669148__fcell-13-1674374-g005.jpg)