hsp47 (MedChemExpress)
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hsp47
Hsp47, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/hy-p2738/Collagens/pmc11149409-428-7-15
Average 93 stars, based on 2 article reviews
Hsp47, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/hy-p2738/Collagens/pmc11149409-428-7-15
Average 93 stars, based on 2 article reviews
hsp47 - by Bioz Stars,
2026-10
93/100 stars
Images
1) Product Images from "Targeting the HSP47-collagen axis inhibits brain metastasis by reversing M2 microglial polarization and restoring anti-tumor immunity"
Article Title: Targeting the HSP47-collagen axis inhibits brain metastasis by reversing M2 microglial polarization and restoring anti-tumor immunity
Journal: Cell Reports Medicine
doi: 10.1016/j.xcrm.2024.101533
Figure Legend Snippet: HSP47 is overexpressed in brain metastasis and high-level HSP47 predicts poor survival in cancer patients (A) Heatmaps showing the differential expression of ECM genes in breast cancer BrMs ( n = 97) compared to primary tumors ( n = 137) (GEO: GSE184869 , GSE12276 , GSE29431 , GSE46928 , and GSE100534 ; GitHub: JNCI_2018) and in lung cancer BrMs ( n = 144) compared to primary tumors ( n = 166) (GEO: GSE14108 and GSE13213 ; OMIX: OMIX575; BioProject: PRJNA681304; ArrayExpress: E_MTAB_8659). (B) Venn diagram showing the overlapping genes that are co-upregulated in BrMs in (A). (C) Diagram showing the in vivo selection procedure of brain metastatic cell populations (BMT5) from 4T1 breast cancer cells and LLC1 lung cancer cells, respectively. (D) 4T1-BMT5 and the parental 4T1 cells (5 × 10 4 cells/mouse) expressing the firefly luciferase were intracardially implanted into immunocompetent BALB/c mice, while LLC1-BMT5 and the parental LLC1 cells (5 × 10 4 cells/mouse) were implanted into C57/BL6 mice, respectively. BrMs were monitored by bioluminescence imaging (BLI) at 28 (4T1 and 4T1-BMT5 cells) or 35 days (LLC1 and LLC1-BMT5 cells) after cell implantation. Representative BLI images are shown (left). BLI photon fluxes in the brain for each mouse were quantified (right, mean ± SD of n = 8 mice, two-sided Mann-Whitney test). ∗∗∗ p < 0.001. (E) Quantification of the percentages of mice that developed bone metastases or BrMs after intracardial implantation of the tumor cells. (F) RT-qPCR assays detected the expression of the 15 co-upregulated genes in 4T1-BMT5 and LLC1-BMT5 cells compared to the parental cells (mean ± SEM of n = 3 independent experiments, two-tailed Student’s t test). GAPDH was used as an internal control. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (G) Immunostaining of HSP47 in BrMs and the paired primary breast (top) or lung cancer tissues (bottom). Representative images of two patients for each cancer type are shown. Scale bars, 100 μm. The expression scores of HSP47 in BrMs were compared with those of the paired primary tumors ( n = 12 for BRCA and n = 26 for LUAD, paired Student’s t test). (H) Kaplan-Meier analysis showing the correlation between HSP47 mRNA levels and overall survival of patients with breast cancer (top) and lung cancer (bottom).
Techniques Used: Expressing, In Vivo, Selection, Luciferase, Imaging, MANN-WHITNEY, Quantitative RT-PCR, Two Tailed Test, Control, Immunostaining
Figure Legend Snippet: HSP47 is required for BrM (A) 4T1 and LLC1 cells expressing HSP47 were analyzed by immunoblotting (top) and RT-qPCR (bottom), respectively. The RT-qPCR data were normalized to GAPDH (mean ± SEM of n = 3 independent experiments, two-tailed Student’s t test). ∗∗∗ p < 0.001. (B) 4T1 and LLC1 cells (5 × 10 4 cells/mouse) expressing HSP47 were intracardially injected into mice, respectively. Representative BLI images of two mice for each group are shown. BLI photon fluxes in the brain for each mouse were quantified (mean ± SD, n = 8 mice for each group, two-sided Mann-Whitney test). ∗∗ p < 0.01. (C) The percentages of mice forming brain and bone metastases after intracardial cell implantation. (D) The BMFS of mice was evaluated ( n = 8 mice for each group, Kaplan-Meier model with two-sided log-rank test). ∗∗ p < 0.01. (E and F) H&E staining showing the BrMs in (B). Representative images are shown (E). Scale bars, 200 μm. Insets: high-magnification images corresponding to the areas marked by dotted red lines. The number of BrMs were quantified (F). Large metastases are >300 μm on the longest axis (mean ± SD, n = 8 mice, two-sided Mann-Whitney test). ∗∗∗ p < 0.001. (G) 4T1-BMT5 and LLC1-BMT5 cells expressing HSP47 shRNAs were analyzed by immunoblotting (top) and RT-qPCR (bottom). For RT-qPCR, GAPDH was used as an internal control. Data are expressed as mean ± SEM, n = 3 independent experiments, two-tailed Student’s t test. ∗∗∗ p < 0.001. (H) 4T1-BMT5 and LLC1-BMT5 cells expressing HSP47 shRNAs were intracardially injected into mice, respectively. Representative BLI images of mice are shown. BLI photon fluxes in the brain for each mouse were quantified (mean ± SD, n = 8 mice for each group, two-sided Mann-Whitney test). ∗∗∗ p < 0.001. (I) The BMFS of mice was evaluated ( n = 8 mice for each group, Kaplan-Meier model with two-sided log rank test). ∗∗∗ p < 0.001. (J) H&E staining of the BrMs in (H). Representative images are shown. Scale bars, 200 μm. Insets: high-magnification images corresponding to the areas marked by dotted red lines. The number of BrMs were quantified (mean ± SD, n = 8 mice, two-sided Mann-Whitney test). ∗∗∗ p < 0.001.
Techniques Used: Expressing, Western Blot, Quantitative RT-PCR, Two Tailed Test, Injection, MANN-WHITNEY, Staining, Control
Figure Legend Snippet: HSP47 promotes M2 microglial polarization and immunosuppression in brain metastatic niche (A) Representative fluorescence-activated cell sorting (FACS) plots and quantification of CD206 + CD11b + CD45 low and CD11c + CD11b + CD45 low cells in the mouse BrMs (mean ± SEM, n = 5 independent experiments, Student’s t tests). ∗∗∗ p < 0.001. (B) Mouse BrMs derived from 4T1-BMT5 cells were double stained with anti-HSP47 and anti-CD206 antibodies. Representative images are shown. Scale bars, 100 μm. The density of CD206 + cells in each microscope filed was counted (mean ± SEM, n = 6 randomly selected fields, Student’s t tests). ∗∗∗ p < 0.001. (C) Representative FACS plots and quantification of CD8 + CD3 + CD45 + cells in the mouse BrMs derived from 4T1-BMT5 cells. Results are presented as mean ± SEM of n = 5 independent experiments, Student’s t tests. ∗∗∗ p < 0.001. (D) LLC1-BMT5 cells expressing HSP47 shRNAs were intracardially injected into mice. Mice were then injected intraperitoneally with a CD8α monoclonal antibody (mAb; 100 μg/mouse) or immunoglobulin G2b (IgG2b) (100 μg/mouse). Representative BLI images and quantification of BLI photon fluxes in the brain are shown (mean ± SD, n = 8 mice for each group, two-sided Mann-Whitney test). ∗∗∗ p < 0.001. (E) Mice intracardially injected with 4T1 or LLC1 cells expressing HSP47 were kept on a standard diet or fed with PLX3397 (290 mg/kg) for 3 weeks. Representative BLI images are shown. BLI photon fluxes in the mouse brain were quantified (mean ± SD of n = 8 mice for each group, two-sided Mann-Whitney test). ∗ p < 0.05 and ∗∗ p < 0.01. (F) H&E staining of BrMs in mice. Scale bars, 200 μm. Large metastases are >300 mm on the longest axis (mean ± SD of n = 8 mice, Student’s t test). ∗ p < 0.05 and ∗∗ p < 0.01. (G) The percentages of mice forming brain or bone metastases were quantified. (H) Representative FACS plots and quantification of CD11b + CD45 low cells in mouse BrMs (mean ± SEM, n = 5 independent experiments, Student’s t tests). ∗∗∗ p < 0.001. (I) Representative FACS plots and quantification of CD8 + CD3 + CD45 + cells in mouse BrMs (mean ± SEM, n = 5 independent experiments, Student’s t tests). ∗∗∗ p < 0.001. (J–M) The populations of Ki67 + , CD69 + , granzyme B (GZMB) + , interferon (IFN)-γ + , and TNF-α + CD8 + T cells and CD25 + FOXP3 + CD4 + Tregs in (I) were quantified (mean ± SEM, n = 5 independent experiments, Student’s t tests), respectively. ∗∗∗ p < 0.001. (N) Immunofluorescence (IF) co-staining of HSP47 and CD206 in brain metastatic lesions of patients with breast cancer. Representative images of two tumors are shown. Scale bars, 100 μm. The expression correlation between HSP47 and CD206 was analyzed ( n = 10 randomly selected fields, Pearson correlation coefficient).
Techniques Used: Fluorescence, FACS, Derivative Assay, Staining, Microscopy, Expressing, Injection, MANN-WHITNEY, Immunofluorescence
Figure Legend Snippet: COL1 mediates HSP47-induced microglia recruitment and polarization (A) ELISA quantification of the COL1 levels in the CM of 4T1-BMT5 and LLC1-BMT5 cells compared to the corresponding parental cells. (B) ELISA quantification of the COL1 levels in the CM of 4T1-BMT5 and LLC1-BMT5 cells expressing HSP47 shRNAs. (A and B) Data are expressed as mean ± SD of n = 3 independent experiments, Student’s t tests. ∗∗ p < 0.01 and ∗∗∗ p < 0.001. (C) BV2 cells were cultured in the CM of 4T1-BMT5 or LLC1-BMT5 cells expressing HSP47 shRNA and then treated with 4 mg/mL COL1 for 24 h. Cells that migrated onto the lower surface of the Transwell chamber were stained by crystal violet. Representative images are shown. Scale bars, 100 μm. Data are presented as mean ± SEM, n = 6 randomly selected microscope fields of two independent experiments, Student’s t tests. ∗∗∗ p < 0.001. (D) BV2 cells cultured in the CM of 4T1-BMT5 or LLC1-BMT5 cells expressing HSP47 shRNA were treated with COL1 for 48 h. The cell lysates were analyzed by immunoblotting using anti-CD206 and anti-Arg1 antibodies. (E) BV2 cells were treated as in (D) and the expression of CD206 was analyzed by FACS. Representative FACS plots are shown. FACS data were statistically analyzed (mean ± SD, n = 3 independent experiments, Student’s t tests). ∗∗ p < 0.01 and ∗∗∗ p < 0.001. (F) 4T1-BMT5 cells expressing HSP47 shRNA were treated with COL1 for 24 h and then intracardially injected into mice. Representative BLI of two mice for each group are shown. BLI photon fluxes in the brain for each mouse were quantified (mean ± SD, n = 8 mice for each group, two-sided Mann-Whitney test). ∗ p < 0.05. (G) The BMFS of mice was evaluated ( n = 8 mice for each group, Kaplan-Meier model with two-sided log-rank test). ∗∗ p < 0.01. (H) 4T1-BMT5 cells expressing control shRNA or HSP47 shRNA were engineered to express GFP and then intracardially injected into mice. Consecutive mouse BrM tissues were immunostained with the antibodies against GFP, HSP47, COL1-4, CD206, or CD8α. Scale bars, 200 μm. Insets: high-magnification images corresponding to the areas marked by dotted red lines. (I) CD8 + T cells were cultured overnight in the CM of tumor cells overexpressing HSP47 or 4 mg/mL COL1. Cells that migrated onto the lower surface of the Transwell chamber were counted by FACS. (J) BV2 cells were treated with the CM of tumor cells expressing HSP47 or 4 mg/mL COL1 overnight. The medium was replaced with fresh medium and then used to treat CD8 + T cells. The T cells that migrated onto the lower surface of the Transwell chamber were counted by FACS. (I and J) Representative FACS plots are shown. FACS data were statistically analyzed (mean ± SD, n = 3 independent experiments, Student’s t tests). ∗ p < 0.05 and ∗∗∗ p < 0.001. (K and L) IF double staining of COL1 and CD206 in human BrMs from patients with breast or lung cancer. Scale bars, 100 μm. The staining density of COL1 and CD206 cells in each tissue was analyzed ( n = 10 tissues for breast cancer BrMs and n = 16 tissues for lung cancer BrMs, Pearson correlation coefficient).
Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, shRNA, Staining, Microscopy, Western Blot, Injection, MANN-WHITNEY, Control, Double Staining
Figure Legend Snippet: HSP47-mediated COL1 biosynthesis promotes M2 microglial polarization through β-integrin/NF-κB signaling (A) BV2 cells were treated with COL1 and then subjected to RNA sequencing analysis ( n = 3 biological replicates for each group). The Kyoto Encyclopedia of Genes and Genomes analysis shows the top 11 enriched pathways with significant difference. (B) Gene set enrichment analysis of the changed genes after COL1 treatment. NES (normalized enrichment score) and p value are shown. (C) Heatmap showing the most significantly upregulated cytokine genes in COL1-treated BV2 cells vs. control BV2 cells. (D) BV2 cells transfected with p65 small interfering RNA (siRNA) were treated with COL1, and the cell lysates were analyzed by immunoblotting. The levels of selected cytokine genes were analyzed by RT-qPCR. (E) BV2 cells under COL1 treatment were treated with TC-I-15. Cell lysates were analyzed by immunoblotting, and the levels of selected cytokine genes were detected by RT-qPCR. (D and E) GAPDH was used as an internal control. Data are expressed as mean ± SEM of n = 3 independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (F) CUT&Tag analysis of p65 binding at CCL2 , CXCL2 , and CXCL3 promoters in BV2 cells after treatment by COL1. Spike-in was used as an internal control. Data are expressed as mean ± SEM, n = 3 independent experiments, two-tailed Student’s t test. ∗∗∗ p < 0.001. (G) BV2 cells under COL1 treatment were transfected with p65 siRNA or treated with TC-I-15 and the expression of CD206 was analyzed by FACS. Representative FACS plots are shown. The FACS data were statistically analyzed (mean ± SEM, n = 3 independent experiments). ∗∗∗ p < 0.001.
Techniques Used: RNA Sequencing Assay, Control, Transfection, Small Interfering RNA, Western Blot, Quantitative RT-PCR, Binding Assay, Two Tailed Test, Expressing
Figure Legend Snippet: Targeting HSP47-mediated COL1 biosynthesis inhibits BrM (A) Molecular formula of Col003. (B) 4T1-BMT5 or LLC1-BMT5 cells were treated with the indicated concentration of Col003 for 24 h and COL1 level in the CM was quantified by ELISA (mean ± SD, n = 3 independent experiments). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. (C) CSF was extracted from mice at different time points after intravenous injection of Col003 (20 mg/kg) and the concentration of Col003 was determined by high-performance liquid chromatography (mean ± SD, n = 3 mice for each time point). (D) 4T1-BMT5 or LLC1-BMT5 cells were intracardially injected into mice. Seven days later, Col003 was injected (5 or 20 mg/kg) into mice via vein every third day for 1 month. Representative BLI images are shown. BLI photon fluxes in the brain for each mouse were quantified (mean ± SD, n = 8 mice for each group, two-sided Mann-Whitney test). ∗∗∗ p < 0.001. (E) H&E staining of BrM tissues in mice. Representative images are shown (left). Scale bars, 200 μm. Large metastases are >300 mm on the longest axis (mean ± SD, n = 8 mice, two-sided Mann-Whitney test). ∗∗ p < 0.01 and ∗∗∗ p < 0.001. (F) The BMFS of mice was evaluated ( n = 8 mice for each group, Kaplan-Meier model with two-sided log-rank test). ∗∗∗ p < 0.001. (G) Representative FACS plots and quantification of CD8 + CD3 + CD45 + cells in the mouse BrM tissues. (H) Representative FACS plots and quantification of CD206 + CD11b + CD45 low and CD11c + CD11b + CD45 low cells in the BrM tissues. (I–L) The populations of Ki-67 + , CD69 + , GZMB + , IFN-γ + , and TNF-α + CD8 + T cells and CD25 + FOXP3 + CD4 + Tregs were analyzed by FACS. (G–L) Data are expressed as mean ± SEM, n = 5 independent experiments, Student’s t tests. ∗∗∗ p < 0.001. (M) Consecutive mouse BrM tissues were analyzed by immunohistochemistry using the antibodies against GFP, COL1, and CD8α. Representative images are shown. Scale bars, 200 μm.
Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay, Injection, High Performance Liquid Chromatography, MANN-WHITNEY, Staining, Immunohistochemistry
Figure Legend Snippet: Col003 augments the therapeutic efficacy of anti-PD-L1 antibody in BrMs (A) 4T1-BMT5 or LLC1-BMT5 cells were intracardially implanted into mice. Seven days after cell injection, mice were intraperitoneally injected with anti-PD-L1 antibody alone (200 μg/mouse) every third day four consecutive times or with anti-PD-L1 plus Col003 (20 mg/kg) (top). Representative BLI images of each group are shown. BLI photon fluxes in the brain for each mouse were quantified (mean ± SD, n = 8 mice for each group, two-sided Mann-Whitney test). ∗∗∗ p < 0.001. (B) The BMFS of mice was evaluated ( n = 8 mice for each group, Kaplan-Meier model with two-sided log-rank test). ∗∗∗ p < 0.001. (C) Representative FACS plots and quantification of CD206 + CD11b + CD45 low and CD11c + CD11b + CD45 low cells in the BrM tissues. (D) Representative FACS plots and quantification of CD45 + CD3 + CD8 + cells in the BrM tissues. (C and D) Data are expressed as mean ± SEM of n = 5 independent experiments, Student’s t tests. ∗∗∗ p < 0.001. (E) Co-staining of GZMB and CD8α in mouse BrM tissues. Representative images are shown (left). Scale bars, 100 μm. The GZMB + and CD8α + positive cells in each microscope field were counted (mean ± SEM, n = 6 randomly selected fields for each group, Student’s t tests). ∗∗∗ p < 0.001. (F) Schematic diagram shows the findings revealed in this study. Upregulation of HSP47 in tumor cells promotes collagen deposition in the brain metastatic niches, which promotes M2 microglial polarization and expression of anti-inflammatory cytokines through the integrin α2β1/NF-κB pathway, resulting in repression of anti-tumor CD8 + T cell immunity.
Techniques Used: Injection, MANN-WHITNEY, Staining, Microscopy, Expressing
Figure Legend Snippet:
Techniques Used: Virus, Recombinant, Control, Lysis, Enzyme-linked Immunosorbent Assay, Isolation, Cell Isolation, Software, Membrane
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