cell lines 5637  (ATCC)

Journal: iScience

Article Title: KNSTRN knockdown impairs autophagy flux to inhibit bladder cancer progression

doi: 10.1016/j.isci.2026.114734

Figure Lengend Snippet: Lysosomal reactivation restores autophagy damage caused by KNSTRN knockdown (A and B) T24 and 5637 cells pretreated with clioquinol (ClioQ, T24, 10 μM; 5637, 20 μM) for 1 h and subsequently transfected with si-NC and si-KNSTRN for 48 h. And subjected to western blot analysis with antibodies against KNSTRN and P62. Representative western blot images and the relative quantification of protein expression are shown. (C) 5637 cell transfected with mCherry-GFP-LC3 were treated with si-NC and KNSTRN-silenced for 48 h in the absence or presence of clioquinol (ClioQ, 20 μM). The number of LC3 puncta (yellow for autophagosomes, red for autolysosomes) was quantified. And images were captured using a fluorescence microscope. Scale bars, 20 μm. (D) Lysosomal acidity was analyzed by LysoTracker red (scale bar: 20 μm). Representative fluorescent images of the cells are shown. The fluorescence intensity is calculated using ImageJ software. Significance is determined by one-way ANOVA. Data are presented as the mean ± SD ( n = 3). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: 5637 cell line was obtained from Servicebio (Wuhan, China).

Techniques: Knockdown, Transfection, Western Blot, Quantitative Proteomics, Expressing, Fluorescence, Microscopy, Software

Journal: iScience

Article Title: KNSTRN knockdown impairs autophagy flux to inhibit bladder cancer progression

doi: 10.1016/j.isci.2026.114734

Figure Lengend Snippet: KNSTRN knockdown-induced ROS is responsible for impairing lysosomal activity and autophagy (A) Reactive oxygen species (ROS) levels were detected through inverted fluorescence microscope in T24 and 5637 cells transfected with si-NC and si-KNSTRN. (B) Lysosomal acidity was analyzed by LysoTracker red (scale bar: 20 μm). Representative fluorescent images of the cells are shown. The fluorescence intensity is calculated using ImageJ software. (C) T24 and 5637 cells pretreated with N-acetylcysteine (NAC, T24, 2 mM; 5637, 5 mM) for 1 h and subsequently transfected with si-NC and si-KNSTRN for 48 h. And subjected to western blot analysis with antibodies against KNSTRN and P62. Representative western blot images and the relative quantification of protein expression are shown. (D) 5637 cell transfected with mCherry-GFP-LC3 were treated with si-NC and KNSTRN-silenced for 48 h in the absence or presence of N-acetylcysteine(NAC;5 mM). The number of LC3 puncta (yellow for autophagosomes, red for autolysosomes) was quantified. Scale bars, 20 μm. Significance is determined by unpaired t test (two groups) and one-way ANOVA (more than two groups). Data are presented as the mean ± SD ( n = 3). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: 5637 cell line was obtained from Servicebio (Wuhan, China).

Techniques: Knockdown, Activity Assay, Fluorescence, Microscopy, Transfection, Software, Western Blot, Quantitative Proteomics, Expressing