Fig 1: Proteasomal degradation of ANXA2 in the knockdown of S100A11. A, Knockdown of S100A11 decreased the expression of ANXA2, while overexpression of S100A11 elevated the expression of ANXA2. B, U87 transfected with shCtrl, sh-S100A11-1 or sh-S100A11-2, and the cells were treated with or without 10 µM of MG132, and the cell lysates were immunoblotted. C, U87 transfected with shCtrl, or sh-S100A11-1 were treated with 50 µg/mL cycloheximide. Whole-cell lysates were harvested at the indicated times. Knockdown S100A11 decreased the half-life of ANXA2. D, The interaction between S100A11 and ANXA2 was confirmed by co-immunoprecipitation in U87 and U251 cells. E, U87 cells were co-transfected with plasmids expressing HA-Ub and Myc-ANXA2 together with either shCtrl or sh-S100A11-1. U251 cells were co-transfected with plasmids expressing HA-Ub and Myc-ANXA2 together with either Vector or S100A11 plasmid. Cells were treated with MG132 for 6 h before cell lysates were immunoprecipitated using a denature IP protocol to pull down ANXA2 protein, and the polyubiquitinated ANXA2 protein was detected by anti-HA antibody
Fig 2: Schematic representation of the protective effects of S100A11 overexpression on neuronal cell apoptosis induced by ischemic stroke.a The stroke-induced decrease in S100A11 expression causes the nuclear accumulation of ANXA1, which ultimately activates the apoptosis pathway. b In comparison, overexpression of exogenous S100A11 promotes ANXA1 membrane translocation and inhibits ANXA1 nuclear translocation by competing with importin ß for NTS sequence of ANXA1, ultimately protecting neuronal cells from stroke-induced apoptosis
Fig 3: Amino acids 42–98 of S100A11 are required to specifically inhibit the nuclear translocation of ANXA1 and decrease the expression of apoptosis-related proteins.a Co-IP showing the interaction of ANXA1 with S100A11 in N2a cells transfected with Flag-S100A11 (WT), (1–41), and (42–98) plasmids. b–e Western blot showing the subcellular localization of ANXA1 in N2a cells transfected with Flag-S100A11 (WT), (1–41), and (42–98) plasmids after OGD/R, and the quantitative analysis of subcellular ANXA1 levels. Data are reported as the means ± S.E.M. from three independent experiments. f, g Images of immunofluorescence staining showing the subcellular localization of ANXA1 in N2a cells transfected with Flag-S100A11 (WT), (1–41), and (42–98) plasmids after OGD/R, and quantitative analysis of nuclear/cytoplasmic ANXA1 levels. Scale bar = 20 µm. h qPCR analysis of Bid mRNA expression in N2a cells transfected with Flag-S100A11 (42–98) and/or GFP-ANXA1 plasmids after OGD/R. i, j Western blots showing the levels of tBid, cleaved PARP and cleaved caspase-3 proteins in N2a cells and statistical analysis of apoptosis-related protein expression. k, l TUNEL staining (k) and MTT assay (l) showing the effect of S100A11 (42–98) on the apoptosis and viability of N2a cells subjected to OGD/R. n.s: the difference between the two groups was not significant; *P < 0.05; **P < 0.01; ***P < 0.001. Data are reported as the means ± S.E.M. from three independent experiments
Fig 4: S100A11 enhances learning and memory function and motor ability in mice following ischemic stroke.a–d Mean escape latency (a) and distance swam (c) to the hidden platform for the sham, MCAO, MCAO + Ad-GFP, and MCAO + Ad-S100A11 groups over 6 days are plotted against the blocks of trials. Mean escape latency (b) and distance swam (d) to the hidden platform on day 6 in the Morris water maze test. *P < 0.05 compared with the sham group; #P < 0.05 compared with the MCAO group; n = 10 mice. e, f Time (in seconds, s) spent in the target quadrant (e), and the number of times the mouse crossed the target platform location (f) during the probe trials on day 7. n = 10 mice. g Representative path tracings on day 7 during the probe trials. h, i The total distance traveled (h) and the time each mouse spent in the center area (i) were analyzed for 10 min during the open field test. n = 10 mice. j The time mice were able to stay on the rotarod. n = 10 mice; data are presented as the means ± S.E.M., *P < 0.05; **P < 0.01
Fig 5: LBX2-AS1 promotes colorectal cancer cell proliferation and invasion via mediating the miR-491-5p/S100A11 axis. (A) The expression of LBX2-AS1 was assessed in HT29 and SW620 cells transfected with pcDNA-LBX2-AS1 via RT-qPCR. **P<0.01 vs. empty vector. (B) The expression of S100A11 mRNA was determined in HT29 and SW620 cells transfected with pcDNA-LBX2-AS1 or si-LBX2-AS1 via RT-qPCR. **P<0.01. (C and D) The expression of S100A11 protein was measured in HT29 and SW620 cells transfected with pcDNA-LBX2-AS1 and/or miR-491-5p mimics according to western blotting. **P<0.01. (E) The cell viability of HT29 and SW620 cells was determined via a Cell Counting Kit-8 assay after transfection with pcDNA-LBX2-AS1 and/or miR-491-5p mimics. **P<0.01 vs. empty vector. (F) Clone formation and (G) invasive ability of HT29 and SW620 cells were assessed after transfection with pcDNA-LBX2-AS1 and/or miR-491-5p mimics. **P<0.01. LBX2-AS1, LBX2 antisense RNA 1; miR, microRNA; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; si-, small interfering RNA.
Supplier Page from Abcam for Anti-S100A11 antibody [EPR11172]