Fig 2: sUmod silencing ameliorates I/R-induced myocardial inflammation and apoptosis. (a and b) Pathological changes and injury score in myocardial I/R injury (HE staining, bar = 100 µm). (c–g) Western blot was used to quantitatively analyze the expression of IL-1ß, IL-6, TNF-a, and IL-10 in myocardial I/R injury. (h–l) Western blot was used to quantitatively analyze the expression of caspase-7, caspase-9, Bax, and Bcl-xL in myocardial I/R injury. *P < 0.05 vs. Sham; #P < 0.05 vs. I/R.

Fig 3: miR-505-5p overexpression attenuated I/R-induced myocardial inflammation and apoptosis. (a and b) The Effects of ov-miR-505-5p and ov-sUmod on myocardial pathological changes and injury score after myocardial I/R injury (HE staining, bar = 100 µm). (c–g) Quantitative analysis of IL-1ß, IL-6, TNF-a, and IL-10 in myocardial I/R injury with ov-miR-505-5p and ov-sUmod by western blot analysis. (h–l) Quantitative analysis of caspase 7, caspase 9, Bax, and Bcl-xL in myocardial I/R injury with ov-miR-505-5p and ov-sUmod by western blot analysis. *P < 0.05 vs. Sham; #P < 0.05 vs. I/R; &P < 0.05 vs. ov-miR-505-5p.

Fig 4: LINP1 functions in repressing UPR and downstream apoptotic genes. A, C qPCR validations of key gene expression in endoplasmic reticulum signaling including GRP78, XBP1, DDIT3 and DR5 in response to LINP1 knockout and overexpression. Each experiment was performed in at least triplicate and results are presented as mean ± s.d. One-way ANOVA and Dunnett’s multiple comparison test were used to analyze the data (*P < 0.05, **P < 0.01, ***P < 0.001). B, D Verifications of key protein expression in endoplasmic reticulum signaling including p-eIF2a, GRP78, XBP1, ATF4, DDIT3 and DR5 in response to LINP1 knockout and overexpression by Western blot. E Trypan blue staining and Sytox Green staining were performed to evaluate the cell death induced by LINP1 depletion and overexpression in cSCC cells. Scale bars, 50 µm. F, G Apoptosis assay by Annexin V-APC/7-AAD double staining was performed in cSCC cells after LINP1 knockdown or overexpression. H TUNEL assay was performed to detect apoptosis after LINP1 knockdown or overexpression. Scale bars, 100 µm. I ProCaspase-8, cleaved Caspase-8, proCaspase-7, cleaved Caspase-7, proCaspase-3 and cleaved Caspase-3 were detected by Western blot in cSCC cells after LINP1 depletion or overexpression. *P < 0.05, **P < 0.01, ***P < 0.001)

Fig 5: SALIS associates with STAT5A to transcriptionally regulate the expression of IGFBP3 and Caspase-7.a Two peptides were identified by high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis of the protein mix precipitated by in vitro-transcribed Biotin-labeled SALIS RNA. b SALIS RNA transcript was used to retrieve STAT5A protein by RNA pull-down assay with beads only and EGFP as controls. The resulting protein mix was applied to detect STAT5A and HK2 by western blot. c RNA immunoprecipitation (RIP) assay was conducted using antibodies against STAT5A and IgG as control. d, e Predicted binding site of STAT5A (diamond) at the promoter regions of IGFBP3 and Caspase-7 by UCSC genome browser (http://genome.ucsc.edu/) and rVista (https://rvista.dcode.org/). f, g The binding enrichment of STAT5A at the above binding sites on the promoter regions of IGFBP3 and Caspase-7 was detected by ChIP-PCR after knockdown of SALIS or STAT5A. h Chromatin isolation by RNA purification (ChIRP) assay with both even and odd antisense oligos tiling SALIS shows the degree and specificity of SALIS retrieval. LacZ tiling oligo retrieval of SALIS RNA was used as a negative control. i, j Relative retrieval of genomic DNAs corresponding to the IGFBP3 or Caspase-7 promoter or GAPDH loci following SALIS or LacZ ChIRP. k Co-localization of SALIS with STAT5A in HepG2 cells was detected using FISH for SALIS and IF staining for STAT5A and observed by confocal microscope. Scale bar: 20 μM. The data are presented as means ± s.d., n = 3. *P < 0.05, **P < 0.01, ***P < 0.001.