Journal: Cell Death & Disease

Article Title: Long noncoding RNA GDIL acts as a scaffold for CHAC1 and XRN2 to promote platinum resistance of colorectal cancer through inhibition of glutathione degradation

doi: 10.1038/s41419-025-07374-w

Figure Lengend Snippet: A Left: complementary sequence analysis between GDIL and CHAC1 mRNA; Right: CHAC1 mRNAs bind with sense, antisense and CHAC1 binding site mutated (MUT CHAC1-site ) GDIL were examined by biotin pull-down assay and qRT-PCR. B The stability of CHAC1 mRNA in SKOV3CR (up) with GDIL knockdown, and SKOV3 (bottom) with wildtype or mutated GDIL overexpression was measured by qRT-PCR. RNA synthesis was blocked by α-amanitin, RNA levels were calculated relative to 0 h. C Venn analysis was performed to identify proteins pulled down by GDIL in both SKOV3 and HCT116 cells by our RNA pull down/Mass spectrometry analysis and also bind with CHAC1 mRNA from ENCODE eCLIP data. D MS2-pull down combined with qRT-PCR and immunoblotting showing interaction between XRN2 and GDIL. E RNA pull-down using GDIL and CHAC1 transcript segments. F RIP showing the interaction between XRN2 domains and GDIL, XRN2 domains and CHAC1 transcript. G Left: relative CHAC1 mRNA binding with XRN2 in control and GDIL silenced cells, represented as the percentage of the input bound; Right: relative CHAC1 mRNA binding with XRN2 in cells transfected with vector, overexpression of wild type and 1–200 nt (fragment 1) deleted GDIL, represented as the percentage of the input bound. H HCT116-CDXs transfected with vector, overexpression of wild type and 1–200 nt (fragment 1) deleted GDIL under oxaliplatin treatment shown in representative images (up), tumor growth (bottom left) and weight change (bottom right).

Article Snippet: Cell lysates were collected and incubated with anti-XRN2 antibody (Cell Signaling Technology Beverly, MA, USA) and Protein G Magnetic Beads (Thermo Fisher Scientific) for 4 h at 4°C.

Techniques: Sequencing, Binding Assay, Pull Down Assay, Quantitative RT-PCR, Knockdown, Over Expression, Mass Spectrometry, Western Blot, Control, Transfection, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: Long noncoding RNA GDIL acts as a scaffold for CHAC1 and XRN2 to promote platinum resistance of colorectal cancer through inhibition of glutathione degradation

doi: 10.1038/s41419-025-07374-w

Figure Lengend Snippet: A Poly(A) + CHAC1 mRNA bind with sense, antisense and CHAC1 binding site mutated (MUT CHAC1-site ) GDIL were examined by biotin pull-down assay and qRT-PCR. B Relative Poly(A) + CHAC1 mRNA binding using anti-XRN2 antibody or IgG in HCT116 cells, represented as the percentage of the input bound. C Stability of Poly(A) + CHAC1 mRNA in SKOV3 and HCT116 cells with control, overexpression of wildtype or fragment 1 deleted GDIL was measured by qRT-PCR. D RNA FISH for GDIL (red) and immunofluorescence (IF) for XRN2 (green) in parental and resistant HCT116 cells. Blue: DAPI; right: merge. Scale bars, 10 mm. E RNA FISH for GDIL (red) and immunofluorescence (IF) for XRN2 (green) in HCT116 cells with GDIL overexpressed. Blue: DAPI; right: merge. Scale bars, 10 mm. F Subcellular localization of XRN2 protein was detected by immunoblotting in HCT116 cells. SNRP70 was used as nuclear control. GAPDH was used as cytoplasmic control. G Schematic model of GDIL/XRN2/CHAC1 binding.

Article Snippet: Cell lysates were collected and incubated with anti-XRN2 antibody (Cell Signaling Technology Beverly, MA, USA) and Protein G Magnetic Beads (Thermo Fisher Scientific) for 4 h at 4°C.

Techniques: Binding Assay, Pull Down Assay, Quantitative RT-PCR, Control, Over Expression, Immunofluorescence, Western Blot

Journal: Cell Death & Disease

Article Title: Long noncoding RNA GDIL acts as a scaffold for CHAC1 and XRN2 to promote platinum resistance of colorectal cancer through inhibition of glutathione degradation

doi: 10.1038/s41419-025-07374-w

Figure Lengend Snippet: Top: regulation of GDIL-glutathione degradation axis by XRN2/CHAC1 in platinum resistant cancer cells. Bottom: combination therapeutic strategy against platinum resistance by adding ASO targeting lncRNA GDIL.

Article Snippet: Cell lysates were collected and incubated with anti-XRN2 antibody (Cell Signaling Technology Beverly, MA, USA) and Protein G Magnetic Beads (Thermo Fisher Scientific) for 4 h at 4°C.

Techniques:

Journal: Cell Death & Disease

Article Title: Long noncoding RNA GDIL acts as a scaffold for CHAC1 and XRN2 to promote platinum resistance of colorectal cancer through inhibition of glutathione degradation

doi: 10.1038/s41419-025-07374-w

Figure Lengend Snippet: A Left: complementary sequence analysis between GDIL and CHAC1 mRNA; Right: CHAC1 mRNAs bind with sense, antisense and CHAC1 binding site mutated (MUT CHAC1-site ) GDIL were examined by biotin pull-down assay and qRT-PCR. B The stability of CHAC1 mRNA in SKOV3CR (up) with GDIL knockdown, and SKOV3 (bottom) with wildtype or mutated GDIL overexpression was measured by qRT-PCR. RNA synthesis was blocked by α-amanitin, RNA levels were calculated relative to 0 h. C Venn analysis was performed to identify proteins pulled down by GDIL in both SKOV3 and HCT116 cells by our RNA pull down/Mass spectrometry analysis and also bind with CHAC1 mRNA from ENCODE eCLIP data. D MS2-pull down combined with qRT-PCR and immunoblotting showing interaction between XRN2 and GDIL. E RNA pull-down using GDIL and CHAC1 transcript segments. F RIP showing the interaction between XRN2 domains and GDIL, XRN2 domains and CHAC1 transcript. G Left: relative CHAC1 mRNA binding with XRN2 in control and GDIL silenced cells, represented as the percentage of the input bound; Right: relative CHAC1 mRNA binding with XRN2 in cells transfected with vector, overexpression of wild type and 1–200 nt (fragment 1) deleted GDIL, represented as the percentage of the input bound. H HCT116-CDXs transfected with vector, overexpression of wild type and 1–200 nt (fragment 1) deleted GDIL under oxaliplatin treatment shown in representative images (up), tumor growth (bottom left) and weight change (bottom right).

Article Snippet: Then XRN2-interacting RNAs were purified by RNeasy mini kit (Qiagen) and detected by qRT-PCR.

Techniques: Sequencing, Binding Assay, Pull Down Assay, Quantitative RT-PCR, Knockdown, Over Expression, Mass Spectrometry, Western Blot, Control, Transfection, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: Long noncoding RNA GDIL acts as a scaffold for CHAC1 and XRN2 to promote platinum resistance of colorectal cancer through inhibition of glutathione degradation

doi: 10.1038/s41419-025-07374-w

Figure Lengend Snippet: A Poly(A) + CHAC1 mRNA bind with sense, antisense and CHAC1 binding site mutated (MUT CHAC1-site ) GDIL were examined by biotin pull-down assay and qRT-PCR. B Relative Poly(A) + CHAC1 mRNA binding using anti-XRN2 antibody or IgG in HCT116 cells, represented as the percentage of the input bound. C Stability of Poly(A) + CHAC1 mRNA in SKOV3 and HCT116 cells with control, overexpression of wildtype or fragment 1 deleted GDIL was measured by qRT-PCR. D RNA FISH for GDIL (red) and immunofluorescence (IF) for XRN2 (green) in parental and resistant HCT116 cells. Blue: DAPI; right: merge. Scale bars, 10 mm. E RNA FISH for GDIL (red) and immunofluorescence (IF) for XRN2 (green) in HCT116 cells with GDIL overexpressed. Blue: DAPI; right: merge. Scale bars, 10 mm. F Subcellular localization of XRN2 protein was detected by immunoblotting in HCT116 cells. SNRP70 was used as nuclear control. GAPDH was used as cytoplasmic control. G Schematic model of GDIL/XRN2/CHAC1 binding.

Article Snippet: Then XRN2-interacting RNAs were purified by RNeasy mini kit (Qiagen) and detected by qRT-PCR.

Techniques: Binding Assay, Pull Down Assay, Quantitative RT-PCR, Control, Over Expression, Immunofluorescence, Western Blot

Journal: Cell Death & Disease

Article Title: Long noncoding RNA GDIL acts as a scaffold for CHAC1 and XRN2 to promote platinum resistance of colorectal cancer through inhibition of glutathione degradation

doi: 10.1038/s41419-025-07374-w

Figure Lengend Snippet: Top: regulation of GDIL-glutathione degradation axis by XRN2/CHAC1 in platinum resistant cancer cells. Bottom: combination therapeutic strategy against platinum resistance by adding ASO targeting lncRNA GDIL.

Article Snippet: Then XRN2-interacting RNAs were purified by RNeasy mini kit (Qiagen) and detected by qRT-PCR.

Techniques: