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Thermo Fisher trypan blue cell viability assay
(A) Schematic of BCL-X exon 2 and position of intron response element (IRE). (B) RNAseq and DESeq2 analysis of GEnCs treated with glucose soup (GS) showed a significant increase in the IL-6 read counts compared to mannitol (MAN) control (n = 5; *p < 0.05). (C) The increase in IL-6 expression in response to GS was confirmed with qRT-PCR, which persisted in the absence of IL-6 from the GS treatment of GEnCs (n = 3; *p < 0.05 vs. mannitol control as assessed by one-way ANOVA and Tukey post-hoc test). (D) Treatment of GEnCs for 6 h with increasing concentrations of IL-6 alone resulted in a dose-dependent increase in BCL-XS/BCL-XL (n = 3; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (E) IL-6 dose-dependently increased the fluorescence green/red (520/590 nm) ratio, representing an increase in <t>cell</t> apoptosis, using the JC-10 <t>assay</t> (n = 4; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (F) IL-6 dose-dependently increased caspase activation (n = 4; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (G) IL-6 dose-dependently decreased cell <t>viability,</t> as assessed with <t>Trypan</t> <t>blue</t> staining, indicating increased cell apoptosis with increasing IL-6 (n = 4; *p < 0.05 as assessed by one-way ANOVA).
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(A) Schematic of BCL-X exon 2 and position of intron response element (IRE). (B) RNAseq and DESeq2 analysis of GEnCs treated with glucose soup (GS) showed a significant increase in the IL-6 read counts compared to mannitol (MAN) control (n = 5; *p < 0.05). (C) The increase in IL-6 expression in response to GS was confirmed with qRT-PCR, which persisted in the absence of IL-6 from the GS treatment of GEnCs (n = 3; *p < 0.05 vs. mannitol control as assessed by one-way ANOVA and Tukey post-hoc test). (D) Treatment of GEnCs for 6 h with increasing concentrations of IL-6 alone resulted in a dose-dependent increase in BCL-XS/BCL-XL (n = 3; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (E) IL-6 dose-dependently increased the fluorescence green/red (520/590 nm) ratio, representing an increase in cell apoptosis, using the JC-10 assay (n = 4; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (F) IL-6 dose-dependently increased caspase activation (n = 4; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (G) IL-6 dose-dependently decreased cell viability, as assessed with Trypan blue staining, indicating increased cell apoptosis with increasing IL-6 (n = 4; *p < 0.05 as assessed by one-way ANOVA).

Journal: bioRxiv

Article Title: The apoptosis gene BCL-X splice isoforms have opposing effects in diabetic kidney disease: potential treatment target and prognostic value

doi: 10.1101/2025.06.06.658276

Figure Lengend Snippet: (A) Schematic of BCL-X exon 2 and position of intron response element (IRE). (B) RNAseq and DESeq2 analysis of GEnCs treated with glucose soup (GS) showed a significant increase in the IL-6 read counts compared to mannitol (MAN) control (n = 5; *p < 0.05). (C) The increase in IL-6 expression in response to GS was confirmed with qRT-PCR, which persisted in the absence of IL-6 from the GS treatment of GEnCs (n = 3; *p < 0.05 vs. mannitol control as assessed by one-way ANOVA and Tukey post-hoc test). (D) Treatment of GEnCs for 6 h with increasing concentrations of IL-6 alone resulted in a dose-dependent increase in BCL-XS/BCL-XL (n = 3; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (E) IL-6 dose-dependently increased the fluorescence green/red (520/590 nm) ratio, representing an increase in cell apoptosis, using the JC-10 assay (n = 4; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (F) IL-6 dose-dependently increased caspase activation (n = 4; *p < 0.05 vs. 0 ng/ml IL-6 as assessed by one-way ANOVA and Tukey post-hoc test). (G) IL-6 dose-dependently decreased cell viability, as assessed with Trypan blue staining, indicating increased cell apoptosis with increasing IL-6 (n = 4; *p < 0.05 as assessed by one-way ANOVA).

Article Snippet: Trypan blue cell viability assay (ThermoFisher Scientific): Trypan blue stain colors dead cells blue, allowing cell viability to be measured.

Techniques: Control, Expressing, Quantitative RT-PCR, Fluorescence, Activation Assay, Staining