ptecs specific atf6 knockdown  (Obio Technology Corp Ltd)

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: Renal warm I/R injury in mice increases the expression of ATF6 (A) RT-qPCR analysis of ATF6 expression in mouse kidney tissues. (B–C) Representative Western blot images and quantification of ATF6 and activated ATF6 (cATF6) expression in mouse kidney tissues subjected to Sham and ischemia 1h/reperfusion 6h (I/R). (D–E) Representative IHC images and quantification of ATF6 expression in kidney tissues (Scale bars, 100 μm and 50 μm). (F–G) ATF6 expression in cultured HK-2 in response to normoxia and hypoxia/reoxygenation (H/R) treatment was determined by RT-qPCR and Western blot. (H) Quantitative statistical analysis of ATF6 protein. Data represent mean ± SDs, and statistical analysis was performed using t test. (∗ p < 0.05). T-vs-C means IR-vs-Sham.

Article Snippet: For PTECs-specific ATF6 knockdown, adeno-associated viruses (AAV)-based delivery system with pAAV9-MCS-miR30shRNA (Atf6)-WPRE or pAAV9-MCS-miR30shRNA (NC)-WPRE (0.5 × 1012 vg, OBiO Technology, Shanghai, China) were injected by tail vein injection to C57BL/6 mice.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Cell Culture

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: ATF6 plays a protective role in renal ischemia-reperfusion injury (A–B) The levels of serum Cr and BUN in mice. (C–D) The levels of serum IL-6 and TNF-α in mice. (E–F) The representative images and kidney injury assessment score of HE staining of the kidney. Ceapin-A7 (10 μM) is an ATF6 inhibitor, and AA147 (10 μM) is an ATF6 agonist. Data represent mean ± SDs, and statistical analysis was performed using one-way ANOVA. (∗ p < 0.05).

Article Snippet: For PTECs-specific ATF6 knockdown, adeno-associated viruses (AAV)-based delivery system with pAAV9-MCS-miR30shRNA (Atf6)-WPRE or pAAV9-MCS-miR30shRNA (NC)-WPRE (0.5 × 1012 vg, OBiO Technology, Shanghai, China) were injected by tail vein injection to C57BL/6 mice.

Techniques: Staining

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: ATF6 plays a protective role in renal ischemia-reperfusion injury (A–B) The levels of serum Cr and BUN in mice. (C–D) The levels of serum IL-6 and TNF-α in mice. (E–F) The representative images and kidney injury assessment score of HE staining of the kidney. (G–H) RT-qPCR analysis of TNF-α and IL-6 expression in cultured HK-2 in response to H/R treatment. (I–J) RT-qPCR analysis of IL-6 and TNF-α expression in cultured HK-2 in response to H/R using siATF6. Data represent mean ± SDs, and statistical analysis was performed using one-way ANOVA. (∗ p < 0.05).

Article Snippet: For PTECs-specific ATF6 knockdown, adeno-associated viruses (AAV)-based delivery system with pAAV9-MCS-miR30shRNA (Atf6)-WPRE or pAAV9-MCS-miR30shRNA (NC)-WPRE (0.5 × 1012 vg, OBiO Technology, Shanghai, China) were injected by tail vein injection to C57BL/6 mice.

Techniques: Staining, Quantitative RT-PCR, Expressing, Cell Culture

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: ATF6 inhibits the activation of the NF-κB pathway (A) Western blot detection of the expression of NF-κB pathway-related proteins in renal tissue; GAPDH was used as a loading control. (B) Quantitative analysis of protein expression, data represent mean ± SDs, and statistical analysis was performed using t test. (∗ p < 0.05). (C) After hypoxia/reoxygenation in the WT group and knockdown group, the expression of NF-κB pathway-related proteins was detected by Western blot, GAPDH was used as a loading control. (D) Quantitative analysis of protein expression. (E) After hypoxia/reoxygenation in the WT group and the ATF6 overexpression group, the expression of NF-κB pathway-related proteins was detected by Western blot, GAPDH was used as a loading control. (F) Quantitative analysis of protein expression. Data represent mean ± SDs, and statistical analysis was performed using one-way ANOVA. (∗ p < 0.05).

Article Snippet: For PTECs-specific ATF6 knockdown, adeno-associated viruses (AAV)-based delivery system with pAAV9-MCS-miR30shRNA (Atf6)-WPRE or pAAV9-MCS-miR30shRNA (NC)-WPRE (0.5 × 1012 vg, OBiO Technology, Shanghai, China) were injected by tail vein injection to C57BL/6 mice.

Techniques: Activation Assay, Western Blot, Expressing, Control, Knockdown, Over Expression

Journal: iScience

Article Title: ATF6 ameliorates renal warm ischemia-reperfusion injury through FHL2-mediated NF-κB signaling pathway

doi: 10.1016/j.isci.2026.115173

Figure Lengend Snippet: The expression of FHL2 is transcriptionally regulated by ATF6 (A) Expression of FHL2 in RNA-seq after using siATF6. (B) RT-qPCR analysis of FHL2 expression in mouse kidney tissues. (C) Representative Western blot images and quantification of FHL2 expression in mouse kidney tissues (GAPDH as the loading control). (D–E) Representative IHC images and quantification of FHL2 expression in kidney tissues (Scale bars, 100 μm). (F–G) FHL2 expression in cultured HK-2 in response to normoxia and hypoxia/reoxygenation (H/R) treatment was determined by Western blot. GAPDH was used as a loading control. (H–I) The expression of FHL2 and ATF6 in HK-2 cells after ATF6 knockdown was examined by Western blot. GAPDH was used as a loading control; (J-K) The expression of FHL2 and ATF6 in HK-2 cell lines after ATF6 overexpression was examined by Western blot. GAPDH was used as a loading control. (L) The input of ATF6 and IgG binding the FHL2 promoters in HK-2 after hypoxia/physoxia treatment by CHIP-qPCR. (M) Luciferase activity of transfected HEK-293T targeting FHL2 and its mutant after H/R using the dual-luciferase reporter assay. (N) The putative ATF6-binding sites in the FHL2 promoter and the nucleotide sequences representing the predicted binding sequences with the red capital letters indicating core binding elements; Data represent mean ± SDs, and statistical analysis was performed using one-way ANOVA. (∗ p < 0.05).

Article Snippet: For PTECs-specific ATF6 knockdown, adeno-associated viruses (AAV)-based delivery system with pAAV9-MCS-miR30shRNA (Atf6)-WPRE or pAAV9-MCS-miR30shRNA (NC)-WPRE (0.5 × 1012 vg, OBiO Technology, Shanghai, China) were injected by tail vein injection to C57BL/6 mice.

Techniques: Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Control, Cell Culture, Knockdown, Over Expression, Binding Assay, ChIP-qPCR, Luciferase, Activity Assay, Transfection, Mutagenesis, Reporter Assay