EMBO/sd-character-level-ner huggingface dataset:
Plasminogen activation imposes a negative feedback on uPAR-mediated cell adhesion to VN.Figure 1A Effect of plasminogen activation on 293/uPAR cells morphology. 293/uPAR cells were seeded on VN and imaged by time-lapse microscopy. Cells were treated with subsequent additions of 10 nM sc-uPA and 30 nM plasminogen (Plg) and representative phase contrast light microscopy images, taken just before and 2 h after addition of plasminogen are shown. The complete time-lapse recording can be found as Movie EV1. Scale bar, 20 µm. B Effect of plasminogen activation on 293/uPAR cells F-actin cytoskeleton. 293/uPAR cells were seeded on VN and treated with 10 nM sc-uPA (upper panel) or with a combination of 10 nM sc-uPA and 30 nM Plg (lower panel). After 2 hours, cells were fixed, permeabilised and stained with phalloidin. Representative images are shown. C Negative feedback between plasminogen activation and cell adhesion to VN. RTCA experiment of 293/uPAR cells seeded in VN-coated E-Plates is shown. Cells were treated with 10 nM sc-uPA, 30 nM plasminogen and 100 nM a2AP. Stippled vertical lines indicate the time-points at which the cells were treated. Each condition was recorded in quadruplicates and the curves represent the average cell index as a function of time. Cell index values are normalized to the cell index measured before the first treatment and the data are from a representative experiment. D The catalytic activity of both uPA and plasmin (Pli) contributes to the negative feedback between plasminogen activation and cell adhesion. RTCA experiment was conducted with 293/uPARWT cells seeded in VN coated E-plates and treated with 10 nM sc-uPA, tc-uPA and GFD, and 30 nM Plg and Pli as shown in the figure. A representative experiment is shown.E The cleavage of uPAR partially contributes to the negative feedback. 293/uPARR83/89A cells were seeded on VN and treated as in panel D. A representative experiment is shown. F uPARR83/89A is not cleaved by uPA and plasmin. 293/uPARWT and 293/uPARR83/89A cells were seeded on FN-coated tissue culture plates and allowed to adhere before treatment with the indicated reagents: 10 nM sc-uPA and tc-uPA, 30 nM Plg and Pli. After 1 h, cells were lysed, resolved by SDS-PAGE and analysed by immunoblotting using monoclonal antibodies recognizing different epitopes in uPAR [51]. R2 binds an epitope in D3 of uPAR and recognizes the full-length receptor (uPAR) as well as the truncated form lacking D1 (D2D3). R3 binds an epitope in D1 and only recognizes full-length uPAR. The blot is from a representative experiment and the mobility of molecular weight standards is shown.
in human serum and the acyl-chain structures.Figure 1(A) Biosynthetic pathway (schematic) of GM3, from ceramide, and to complex gangliosides. (B) TLC analysis of ganglioside species in human serum.(D) Detailed structures of GM3 species: sialyllactose head group, sphingoid base (d18:1), typical fatty-acid lengths (LCFA, VLCFA), and acyl chain modifications (α-hydroxylation, ω-9 desaturation). (E) Quantification by LC-MS/MS analysis of serum GM3 species with differing acyl chain structures. Total abundance of 10 species was defined as 1. Data expressed as mean ± SD, n = 6.
a) Enhancement of etoposide-induced non-apoptotic death by overexpression of human Bcl-2 and Bcl-xL in Bax/Bak−/− MEFs in a Beclin 1-dependent manner.figf5(a) Enhancement of etoposide-induced non-apoptotic death by overexpression of human Bcl-2 and Bcl-xL in Bax/Bak−/− MEFs in a Beclin 1-dependent manner. Bax/Bak−/− MEFs with silencing of control Bax or Beclin 1 were transfected with the indicated plasmids (1 μg). After 24 h, the transfected cells were incubated with 20 μM etoposide for 24 h, and cell viability was assessed by the CTB assay. Data are shown as mean ± s.d. (n = 3). (b) Bax/Bak−/− MEFs were transiently transfected with 10 μg of siRNA for Bcl-2, Bcl-x, or Bax (negative control) for 24 h. Expression levels of Bcl-2, Bcl-xL and Lamin B (loading control) were analysed by Western blot analysis. (c-e) Inhibition of etoposide-induced autophagy in Bax/Bak−/− cells treated with siRNA for Bcl-x. (c) Bax/Bak−/− MEFs that were transfected with the indicated siRNAs were treated with 20 μM etoposide for 24 h, then examined by phase-contrast microscopy. (d) Bax/Bak−/− MEFs that were transfected with siRNA for Bcl-x and then treated with etoposide for 18 h were analysed by electron microscopy (×8,500) (e) Reduction of punctate GFP-LC3 fluorescence in etoposide-treated Bax/Bak−/− MEFs by silencing of Bcl-x. Bax/Bak−/− MEFs that were transfected with both GFP-LC3 and the indicated siRNAs were incubated without (NT) or with etoposide for 24 h, and then were examined by confocal fluorescent microscopy. The percentage of cells with punctate GFP-LC3 fluorescence was calculated relative to all GFP-LC3-positive cells. n = 4. f) Inhibition of etoposide-induced cell death by silencing of Bcl-x. Bax/Bak−/− MEFs were transfected with siRNA (circles, siRNA for Bax; squares, siRNA for Bcl-x) were treated with 20 μM etoposide in the absence (open symbols) or presence (closed symbols) of 10 mM 3-MA, and cell viability was assessed by the CTB assay at the indicated times. n = 4. (g) Clonogenicity assay of Bax/Bak−/− MEFs after exposure to etoposide and staurosporine. Bax/Bak−/− MEFs that were transfected with the indicated siRNAs were treated with 20 μM etoposide or 1 μM staurosporine at 24 h. After collection, 2,000 cells were seeded in the normal medium. After one week, colonies were counted. h, i) Inhibition of APG5-APG12 induction by silencing of Bcl-x or Beclin 1. Bax/Bak−/− MEFs were treated with the indicated siRNAs (10 μg) for 24 h and then incubated with 20 μM etoposide for the indicated times. Expression of APG5-APG12 complex, Beclin 1 and GAPDH/VDAC (loading control) was analysed by Western blot analysis.
Figure 2(A) Ensemble of 15 lowest-energy structures for Bax (α2-α5) dimer in bicelles calculated from the geometry constraints obtained from the NMR experiments summarized in Table 1. The backbone structures are shown as thin ribbons. (B) Comparison of the NMR structure of Bax (α2-α5) in bicelles (left; PDB code: 6L8V) and the crystal structure (right; PDB code: 4BDU). The NMR or crystal structure has a flat or concave bottom surface formed by the α4-α5 regions, respectively, as indicated by the complimentary flat or convex shapes underneath. The zoom-in regions display the distances between the amide protons of D68-R109' and S60-F100', which are longer in the NMR structure than the crystal structure.
Figure 5a. The activity of Ubc1 was compared with the K48-linked chain building enzyme Ubc7/Cue1 in single turnover ubiquitination experiments in presence of either monoubiquitin, K48‑Ub2 or K63-Ub2. Initial rates were determined b. The K63-linked chain building enzyme Ubc13 with its cofactor Uev1a or the Ubc1 homologue Ube2K (h. sapiens) were employed in single turnover ubiquitination experiments in presence of K63-Ub2 with Ub(K48R) at the proximal or distal position or in presence of K48‑Ub2. Initial rate determination c. Experiment as in (B) but with the Ubc1 homologue ubc-20 (c. elegans).
SchemeTable 1.Figure 7  Figure 7: Scheme Table 1. Summary of selected macrophage RNA-Seq and miRNA data generated in this study. Key pathways and key genes differentially regulated in different groups and conditions. Full datasets have been uploaded as supplementary tables. NOD-like signalling; inflammatory responses
Figure 3A, After 1 day in 5 % LPDS, NPC1-GFP expressing cells were loaded with 50 μg/ml LDL with or without 10 μM PF228 for the indicated times, and stained for intracellular D4H. Representative confocal images of intracellular D4H, NPC1-GFP and internalized integrin β1 labeling. Dashed lines indicate cell outlines. B, Fraction of D4H positive area in the cell as in A. C, Fraction of D4H area overlapping with NPC1 in a peripheral region toward the leading edge (exemplary box shown as inset in A). D, Quantification of D4H area positive for integrin β1 in a peripheral region toward the leading edge. Mean ± SD, n = 26-30 cells pooled from 2 independent experiments. Student's t-test. E-G, Control and FAK siRNA treated cells were LDL depleted for 1 day, loaded with LDL for the indicated times, and stained for plasma membrane D4H or for intracellular D4H with lamp1 or integrin β1 antibodies. Quantification of plasma membrane D4H (E) and intracellular D4H area positive for lamp1 (F) or integrin β1 (G); corresponding images in Fig EV3D, E. Mean ± SD, n = 20-35 cells from 2 independent experiments. Student's t-test. H, Schematic of FAK-regulated PI(4,5) P2 generation on endosomes via PIPKIγ, modified from (Nader et al., 2016). I, After 1 day 5 % LPDS, cells were loaded with 50 μg/ml LDL with or without 10 μM PF228 for the indicated times and endomembrane PI(4,5) P2 was stained with antibody and imaged by confocal microscopy. J, Quantification of mean PI(4,5) P2 immunoreactivity in cells. Mean ± SD, n = 16-26 cells pooled from 2 independent experiments. Student's t-test.
Figure 1(A) HTM of EdU levels per individual nucleus in response to USP7 inhibition in HCT-116 cells treated with 50 μM P22077 for 1-2 hrs or 25 μM BAY 11-7082 for 1-2 hrs, or with DMSO as a control. (B) Western Blot (WB) showing the levels of USP7, MPM-2, histone H3S10P and histone H3 in whole nuclear extracts of HCT-116 cells treated with 50 μM P22077 for 2-4 hrs (USP7i), 10 μM RO3306 for 8 hrs (CDK1i) or DMSO as a control. Ponceau staining is shown as a loading control. (C) Flow cytometry profile illustrating the levels of histone H3S10P (left) and MPM-2 (right) in HCT-116 cells in control conditions (Control, black) and after treatment with 50 μM P22077 for 4 hrs (USP7i, red). DNA content was measured with Propidium Iodide (PI). (D, E) HTM-mediated quantification of MPM-2 (D) and H3S10P (E) levels per individual nucleus in U2OS cells treated with DMSO as a control or 50 μM P22077 (USP7i) for 4 hrs. Nuclei were stained with DAPI (blue). Scale bar, 30 μm. The quantification is shown to the right of representative images for each analysis. (*** p<0.05; t-test). Grey lines indicate mean values. (F) Flow cytometry profile representing the levels of histone H3S10P and MPM-2 in HCT-116 cells transfected with two different siRNA against USP7 (red) (si#1 and si#2) or with a non-specific siRNA (Control, black), for 96h. DNA content was measured with Propidium Iodide (PI). (G) WB illustrating the depletion of USP7 in the HCT-116 cells used in (F). Experiments were repeated 3 times and one representative result is shown.
Figure 1(A) Representative micrographs of optic nerve sections collected from mice 8 days after ONC, showing the accumulation of IB-4+ activated myeloid cells in the injury site, which is demarcated by GFAP staining. Scale bar, 50 µm. (B) Representative micrograph of the lesion site in optic nerve sections 7 days after crush injury in [Cx3cr1gfp/+ WT] BM chimeric mice, immunostained for GFP (red), to depict monocyte-derived macrophages, and IB-4 (green) to label activated myeloid cells. Scale bar, 100 µm. Inset scale, 20 µm. (C) Representative micrograph of optic nerve section 7 days after crush injury, immunostained for CD3 (red, arrows). Scale bar, 100 µm. (D) Representative dotplots showing the general gating strategy used in flow cytometry experiments for the analysis of immune cells in the retina. Leukocytes were identified by gating on CD45+ cells, and were further gated on CD11b+ or TCRβ+ to detect monocytes/macrophages and T cells, respectively. Of the CD11b+ cells, infiltrating monocytes were regarded as CD45hi. (E) Representative contour plot from [Cx3cr1gfp/+ WT] BM chimeric mouse, verifying that infiltrating blood-derived macrophages (CX3CR1-GFP+) are indeed CD45hi, and showing that they are detectable only in retinas from eyes that underwent ONC, while absent in noninjured controls (red vs. blue line). Cells were pre-gated on CD11b. n = 3-4 per group. (F) Quantitative analysis of flow cytometry data, showing the induction of an immune response in the retina after ONC, evident by a general increase in leukocytes (CD45+), including CD11b+CD45hi cells (representing infiltrating macrophages) and T cells (TCRβ+). n = 4 per group. Data shown are representative of six independent experiments. Graphs show mean ± SE of each group. *** P < 0.001. FSC - forward scatter; noninj - noninjured control; ONC - optic nerve crush.
Figure 5(A) CD11c+ cells, CD4+ cells and CX3CR1+CD45intIAb+ (microglia) cells were isolated from peak EAE animals from the CNS. Samples from 6-11 animals were pooled in each experiment; at least two experiments were performed for each subset. Depicted are normalized mRNA expression (to expression in spleen CD11c+ cells) levels as determined by qRT-PCR analysis relative to the housekeeping genes Eukaryotic translation elongation factor 1 alpha 1 (Eef1a1) and Peptidylprolyl isomerase A (Ppia). CNS CD11c+ cells show gene expression of the inflammatory chemokines Ccl2, Ccl5, Cxcl9, Cxcl10 but not Il23 (mean ± SEM).(C) Quantitative expression of EAE-relevant factors Il23 and Csf2 were investigated in the CNS samples as described above (B) CD11c+ cells, CD4+ cells and CD11c-CD11b+IAb+ (monocytes/macrophages) were isolated from EAE affected mice as described in (A). Samples from 6-11 animals were pooled in each experiment; at least two experiments were performed for each subset. Depicted are normalized mRNA expression (to expression in spleenCD11c+ cells) levels as determined by qRT-PCR analysis relative to the housekeeping genes Eukaryotic translation elongation factor 1 alpha 1 (Eef1a1) and Peptidylprolyl isomerase A (Ppia). Relevantly regulated chemokines in other cell populations than CNS CD11c+ cells are shown (mean ± SEM).(D) Quantitative expression of EAE-relevant factors Il23 and Csf2 were investigated in the spleen samples. Figure 5Chemokine expression profiling of CD11c+ cells in the spleen and CNS reveal a key role of CNSCD11c+ cells for inflammatory chemokines.(A) CD11c+ cells, CD4+ cells and CX3CR1+CD45intIAb+ (microglia) cells were isolated from peak EAE animals from the CNS. Samples from 6-11 animals were pooled in each experiment; at least two experiments were performed for each subset. Depicted are normalized mRNA expression (to expression in spleenCD11c+ cells) levels as determined by qRT-PCR analysis relative to the housekeeping genes Eukaryotic translation elongation factor 1 alpha 1 (Eef1a1) and Peptidylprolyl isomerase A (Ppia). CNSCD11c+ cells show gene expression of the inflammatory chemokines Ccl2, Ccl5, Cxcl9, Cxcl10 but not Ccl20 (mean ± SEM).(B) CD11c+ cells, CD4+ cells and CD11c-CD11b+IAb+ (monocytes/macrophages) were isolated from EAE affected mice as described in (A). Relevantly regulated chemokines in other cell populations than CNSCD11c+ cells are shown (mean ± SEM).(C) Quantitative expression of EAE-relevant factors Il23 and Csf2 were investigated in the CNS samples as described above and (D) the spleen samples.Statistical significance was determined using Kruskal-Wallis test. P-values < 0.05 were regarded as statistically significant. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Atg36 is not required for the Cvt pathway, mitophagy or non‐specific autophagy.Figure 2(A) Cvt pathway activity was assessed by endogenous expression of Ape1-GFP in WT, atg36Δ and atg1Δ cells. Cells were grown for 18 h in glucose medium and vacuoles were stained with FM4‐64. Accumulation of GFP in the vacuole indicates normal function of pathway. Bar, 5 μm. (B) Mitophagy of WT, atg36Δ and atg1Δ cells was assayed by western blot analysis of OM45-GFP after culturing cells for up to 3 days in glycerol medium. Samples were taken at 24 h intervals. (C) WT, atg36Δ and atg1Δ cells were assayed for non‐specific autophagy by alkaline phosphatase assay. Cells were grown in YPD and shifted to SD‐N medium for 4 h. Samples were collected and processed for Pho8Δ60 activity. The results represent the mean and s.d. of three experiments. WT 4 h starvation is set at 100%. (A) Cvt pathway activity was assessed by endogenous expression of Ape1-GFP in WT, atg36Δ and atg1Δ cells. Cells were grown for 18 h in glucose medium and vacuoles were stained with FM4‐64. Accumulation of GFP in the vacuole indicates normal function of pathway. Bar, 5 μm.
Figure 4A-C Slices showing lateral association between two plates and the corresponding 3D segmentations. D. Edge-to-edge interactions form cylindrical structures. Data information: Scale bars: 50 nm.
Ccq1 maintains nucleosome stability through recruiting CLRC and SHREC.Figure 3A Scheme of shelterin, CLRC and SHREC complexes and Swi6HP1 bound to H3K9me2 ChIP-qPCR analyses of H3 in WT and mutants as indicated (n = 3 independent experiments). Data information data are represented as mean ± SE ChIP-qPCR analyses of H3 in WT and mutants as indicated (n = 3 independent experiments). Data information data are represented as mean ± SE ChIP-qPCR analyses of H3K9me2 in WT and mutants as indicated (n = 3 independent experiments). Data information data are represented as mean ± SE
TAG .Figure 2(A-D) 25HC inhibits entry of SARS-CoV-2 pseudovirus (A and B), SARS-CoV pseudovirus (C), and MERS-CoV pseudovirus (D). Calu-3 cells were treated with EtOH or indicated concentrations of 25HC for 16h. Cells were then challenged with FLuc-expressing pseudovirus (A, C and D) and EGFP-expressing pseudovirus (B, C and D) at MOI=0.2, with EtOH or 25HC. After 2 h of infection, virus was removed and pseudovirus infection was quantified by luciferase assay at 24 h post infection (A, C and D) or visualized by fluorescence microscopy (B, C and D). IC50 was calculated by Graphpad using non-linear regression analysis. (E) 25HC does not inhibit SARS-CoV-2 pseudovirus receptor binding. Calu-3 cells were treated with EtOH or 25HC (2.5 or 5 µM) for 24h prior to challenge by SARS-CoV-2 pseudovirus at 4 °C for 1h. Total cellular RNA was extracted and quantified for VSV-N by qPCR. 25HC inhibits membrane fusion mediated by SARS-CoV-2 S (F) 293FT or Vero cells were treated with EtOH or 25HC (5 µM) for 16h prior to transfection with pLVX plasmids encoding SARS-CoV-2 S (F) in the absence of EtOH or 25HC. At 24h post-transfection, syncytium formation was visualized by fluorescence microscopy. White arrows indicate syncytia. Quantification of membrane fusion was performed by calculating the percentages of nuclei involved in syncytia formation from all nuclei in GFP positive cells. 25HC inhibits membrane fusion mediated by , SARS-CoV S (G, left), and MERS-CoV S (G, right). 293FT or Vero cells were treated with EtOH or 25HC (5 µM) for 16h prior to transfection with pLVX plasmids encoding , SARS-CoV S (G, left), and MERS-CoV S (G, right) in the absence of EtOH or 25HC. At 24h post-transfection, syncytium formation was visualized by fluorescence microscopy. Scale bar, 50 μm. White arrows indicate syncytia. Quantification of membrane fusion was performed by calculating the percentages of nuclei involved in syncytia formation from all nuclei in GFP positive cells.
is involved in the peroxisome-mediated cholesterol supply into the ciliary membrane (A) Schematic of ORP3 structure.Figure 5Schematic of ORP3 structure. ORP3 contains an N-terminal pleckstrin homology (PH) domain that interacts with phospholipids, an FFAT motif that specifically interacts with VAP-A/B, and a highly conserved C-terminal OSBP-homology domain (OHD). Western blot analysis showing depletion of ORP3 in the ORP3−/− hTERT-RPE1 cell clones. GAPDH served as a loading control. ORP3+/+ and ORP3−/− hTERT-RPE1 cells incubated for 24 h without serum were immunostained with anti-pericentrin (red) and anti-acetylated tubulin (blue) antibodies. Cholesterol was stained with Filipin III (green). Arrows indicate primary cilia. Scale bar, 5 μm. Quantification of the Filipin III intensity at primary cilia from (C). ORP3−/− hTERT-RPE1 cells exhibited a significant reduction of ciliary cholesterol (***p<0.001: one-way ANOVA with Tukey's HSD, n=3: 40-50 cells per experiment). In the boxplot, medians, 25th/75th percentile and min/max were represented by the central lines, the box limits, and the whiskers/error bars, representatively. Three-dimensional reconstitution of the quiescent G0-phase ORP3+/+ and ORP3−/− hTERT-RPE1 cells immunostained with anti-ARL13B (blue), anti-phospho-S473-Akt (red), anti-ORP3 (green), and anti-PMP70 (white) antibodies indicates that ORP3 at the ciliary pocket (arrow and arrowhead) mediates the membrane regions of ciliary pocket and peroxisomes (arrowhead). The scale bars indicate 5 μm. Quantification of proportion of primary cilia interacting with peroxisomes from (E). Depletion of ORP3 significantly interfered with the spatial interaction between peroxisomes and primary cilia (mean ± s.d.: ***p<0.001: one-way ANOVA with Tukey's HSD, n=3: 45-50 cells per experiment). 3×FLAG-tagged ORP3 and AcGFP1-tagged PEX14, EHD1 or EHD3 were coexpressed in HEK293T cells and then immunoprecipitated from whole-cell lysates using the anti-FLAG antibody. 3×FLAG-tagged ORP3 and AcGFP1-tagged PEX14, EHD1 or EHD3 fragments in the IP fractions and inputs were detected by western blotting. ORP3−/− hTERT-RPE1 cells were transfected with AcGFP1-tagged ORP3, PH domain deleted-ORP3 mutant (ΔPH), FFAT motif-deleted-ORP3 mutant (ΔFFAT), or OHD domain deleted-ORP3 mutant (ΔOHD) and cultured without serum for 24 h before Filipin-III (green)-mediated cholesterol staining and immunostaining with anti-GFP (red), anti-pericentrin (white), and anti-acetylated-tubulin (blue) antibodies. Arrows represent AcGFP1-tagged ORP3 or the mutants localized to the ciliary pocket. AcGFP1-tagged ORP3ΔPH mutant mis-localized to the ciliary pocket. The scale bars indicate 5 μm. Quantification of the Filipin III intensity at primary cilia from (H). AcGFP1-tagged ORP3 deletion mutants did not restore the ciliary cholesterol insufficiency in the ORP3−/− hTERT-RPE1 cells (*p<0.05, ***p<0.001: one-way ANOVA with Tukey's HSD, n=3: 40-50 cells per experiment). In the boxplot, medians, 25th/75th percentile and min/max were represented by the central lines, the box limits, and the whiskers/error bars, representatively.
Hsp70 augments inflammation through Siglec‐14 in THP1 cellsSecretion of TNFα by THP1 cells transfected with a Siglec‐5 or Siglec‐14 expression vector was evaluated by ELISA after exposure to Hsp70 at the various concentrations in the presence of endotoxin‐neutralizing agent polymyxin B.Figure 6Secretion of TNFα by THP1 cells transfected with a Siglec‐5 or Siglec‐14 expression vector was evaluated by ELISA after exposure to Hsp70 at the various concentrations in the presence of endotoxin‐neutralizing agent polymyxin B. Only THP1 Siglec‐14 cells exhibited significant secretion of TNFα in an Hsp70 dose‐dependent manner. IL‐8 secretion was also evaluated after THP1Siglec‐5 and Siglec‐14cells were exposed to Hsp70. Similarly, only THP1Siglec‐14cells exhibited significant secretion of IL‐8 in an Hsp70 dose‐dependent manner. Western blot analysis demonstrates a greater increase in phosphorylated p38 (p‐p38) from THP1 Siglec‐14 cells after exposure to Hsp70 in comparison with control THP1 EV cells in a time‐dependent manner. Graph below immunoblots indicates densitometric analysis of each band normalized to the respective cell line's unstimulated control group, divided by the respective loading control. Exposure to Hsp70 also yielded greater degradation of the NF‐κB inhibitor, IκBα, in THP1 Siglec‐14 cells in comparison with control THP1 cells, augmentation of the pro‐inflammatory response. Graph below immunoblots indicates densitometric analysis of each band normalized to the respective cell line's unstimulated control group, divided by the respective loading control. Secretion of pro‐inflammatory cytokine IL‐8 by PMA‐differentiated THP1 EV or THP Sig 14 cells was evaluated by ELISA after concurrent exposure with an anti‐Hsp70 antibody or isotype‐matched antibody. Secretion of IL‐8 was decreased in only THP1 Sig 14 cells exposed to LPS and an anti‐Hsp70 antibody in comparison with the isotype control antibody, but this change was not observed in THP1 EV cells.