Journal: Apoptosis

Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

doi: 10.1007/s10495-026-02262-w

Figure Lengend Snippet: NHE7 activates ER stress pathways in vitro and in vivo. A WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 overexpressing. B WB analysis of ER stress markers (p-PERK, p-IRE1α, ATF6) in Ishikawa, HEC-1-A, and HEC-1-B cells following NHE7 knockdown. C IHC staining of p-IRE1α and ATF6 in xenograft tumor tissues with NHE7 overexpression. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). D Correlation analysis between NHE7 and ER stress markers (p-IRE1α, ATF6) in clinical EC samples by IHC. Scale bars, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

Techniques: In Vitro, In Vivo, Knockdown, Immunohistochemistry, Over Expression

Journal: Apoptosis

Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

doi: 10.1007/s10495-026-02262-w

Figure Lengend Snippet: NHE7 enhances malignant phenotypes and stemness in EC cells by activating the ER stress pathway. A MTT assays were performed to evaluate the effect of 4-PBA concentration on cell viability in Ishikawa cells. Ishikawa cells were divided into pCDH + Control, NHE7 + control and NHE7 + 4-PBA (ER stress inhibitor). B The expressions of p-IRE1α, IRE1α and ATF6 were examined in Ishikawa cells through western blot assay. C The proliferation ability of Ishikawa cells was detected using the MTT assay. D The apoptosis levels of Ishikawa cells were detected using flow cytometry. E The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. F The colony-forming capacity of Ishikawa cells was tested using a colony formation assay. G The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bars, 100 μm (100 ×). H Tumor sphere-forming assay showed the effects of NHE7 overexpression and 4-PBA on the tumor sphere formation of Ishikawa cells. I WB analysis assessed the expression levels of EMT markers (E-cadherin, N-cadherin, and Vimentin) and stemness-associated proteins (OCT4, Nanog, and SOX2). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05, ** P < 0.01

Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

Techniques: Concentration Assay, Control, Western Blot, MTT Assay, Flow Cytometry, Colony Assay, Migration, Over Expression, Expressing

Journal: Apoptosis

Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

doi: 10.1007/s10495-026-02262-w

Figure Lengend Snippet: NHE7 enhances OXPHOS-induced ER stress by upregulating COX6C expression in EC cells. A WB was used to detect the transfection efficiency of COX6C knockdown in Ishikawa cells. B WB was used to analyze the expression of NHE7 and COX6C in Ishikawa cells after COX6C silencing combined with NHE7 overexpression. C ELISA was used to determine the level of ATP. D Intracellular ROS levels were detected by flow cytometry. E The expression levels of p-IRE1α, IRE1α, and ATF6 were examined in Ishikawa cells. F The apoptosis levels of Ishikawa cells were detected using flow cytometry. G The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. H The colony-forming capacity of Ishikawa cells was tested by colony formation assay. I The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bar, 100 μm (100 ×). J Tumor sphere-forming assay showed the effects on the tumor sphere formation of Ishikawa cells. At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

Techniques: Expressing, Transfection, Knockdown, Over Expression, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Colony Assay, Migration

Journal: Apoptosis

Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

doi: 10.1007/s10495-026-02262-w

Figure Lengend Snippet: COX6C triggers ER stress and enhances the malignant phenotypes of EC cells by upregulating ROS levels. A Intracellular ROS levels were detected by flow cytometry in Ishikawa cells with COX6C overexpression combined with the antioxidant N-acetylcysteine (NAC). B The expression levels of p-IRE1α, IRE1α, and ATF6 were examined in Ishikawa cells. C The proliferation ability of Ishikawa cells was detected using the MTT assay. D The apoptosis levels of Ishikawa cells were detected using flow cytometry. E The expressions of apoptosis-related markers (cleaved PARP and cleaved Caspase-3) in Ishikawa cells were detected by WB assay. F The colony-forming capacity of Ishikawa cells was tested by colony formation assay. G The migration and invasion abilities of Ishikawa cells were detected through transwell experiments. Scale bar, 100 μm (100 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

Techniques: Flow Cytometry, Over Expression, Expressing, MTT Assay, Colony Assay, Migration

Journal: Apoptosis

Article Title: Hypoxic glycolysis-driven histone lactylation activates NHE7 to promote endometrial cancer progression via COX6C-mediated endoplasmic reticulum stress

doi: 10.1007/s10495-026-02262-w

Figure Lengend Snippet: NHE7 facilitates EC tumor growth by upregulating COX6C expression in vivo. A WB analysis was used to detect the protein expression levels of NHE7 and COX6C in Ishikawa cells overexpressing NHE7, and in Ishikawa cells with concurrent NHE7 overexpression and COX6C knockdown. B Stable Ishikawa cell lines were established for the following conditions: NHE7 overexpression alone, and concurrent NHE7 overexpression with COX6C knockdown, along with their respective control cells. A xenograft tumor model was then generated using these cells. C Tumor growth was monitored and recorded, and D tumor weight was measured at the endpoint. E WB assay was performed to assess the effects of NHE7 overexpression alone or in combination with COX6C knockdown on the levels of apoptosis markers c-PARP and C-Caspase3 in xenograft tumor tissues. F HE staining was performed to assess histopathological damage in xenograft tumor tissues following NHE7 overexpression, either alone or in combination with COX6C knockdown. IHC was performed to evaluate the impact of NHE7 overexpression, either alone or in combination with COX6C knockdown, on the expression of Ki67, E-cadherin, N-cadherin, Vimentin, OCT4, Nanog, SOX2, p-IRE1α, IRE1α, and ATF6 in xenograft tumor tissues. Scale bar, 50 μm (400 ×) and 100 μm (200 ×). At least three independent experiments were performed on the assays. Error bars represent the mean ± SD ( n = 3). * P < 0.05,** P < 0.01

Article Snippet: p-IRE1α , BIOSS , bs-16698R , WB/IHC.

Techniques: Expressing, In Vivo, Over Expression, Knockdown, Control, Generated, Staining

Journal: iScience

Article Title: SEL1L confers reduced EMT and drug resistance to breast cancer cells by promoting Shh degradation

doi: 10.1016/j.isci.2025.113876

Figure Lengend Snippet: SEL1L promoted Shh protein degradation via affecting the ubiquitination-proteasome system in TNBC cells (A) SEL1L-induced reduction of Shh protein was rescued in the present of the proteasome inhibitor MG132 in 231 CR cells. (B) Western blot results showed the Shh protein elevated by SEL1L shRNA can be further enhanced via MG132. (C and D) The half-life of Shh protein was shortened by SEL1L overexpression in the presence of the protein synthesis inhibitor CHX in 231 CR) and 549 CR cell. (E) Western blot results showed SEL1L overexpression promoted ubiquitination of Shh protein in 231 CR cells. (F) Silencing of SEL1L significantly decreased ubiquitination of Shh protein in 231 cells. (G) Western blot analysis was performed to assess the expression levels of HRD1 in M-231cells following SEL1L knockdown. (H) The expression of HRD1 was downregulated by three HRD1 shRNA in M-231 cells. (I) Western blot results showed silenced HRD1 decreased ubiquitination of Shh protein in M-231 cells. (J) In SEL1L-overexpressed 231 CR cells, HRD1 knockdown rescued the increased ubiquitination of Shh protein. (K) In SEL1L-overexpressed 231 CR cells, following HRD1-C2A mutation, the SEL1L overexpression-induced enhancement of Shh ubiquitination was effectively reversed. (L) M-231 NC or SEL1L shRNA cells were fractionated into two parts (whole cell and endoplasmic reticulum (ER)) and blotted with antibodies to Erp27 (ER marker), and tubulin (cytosol marker). SEL1L levels was downregulated both in WCL and ER, Shh protein was accumulated within the ER following SEL1L knockdown. (M) Western blot analysis was performed to assess the expression levels of IRE1α in M-231 and BT549 cells following SEL1L knockdown. Data are shown as mean ± SD of three independent replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: IRE1α Antibody , Proteintech , Cat#27528-1-AP; RRID: AB_2880899.

Techniques: Ubiquitin Proteomics, Western Blot, shRNA, Over Expression, Expressing, Knockdown, Mutagenesis, Marker