Journal: Mucosal Immunology

Article Title: Butyrate regulates neutrophil homeostasis and impairs early antimicrobial activity in the lung

doi: 10.1016/j.mucimm.2023.05.005

Figure Lengend Snippet: Effect of butyrate on acute Pseudomonas aeruginosa immunity. (A) Quantification of the total number of cells in BALF 18 hours post-inoculation with 1 × 10 6 CFU of P. aeruginosa strain PAO1 in Ctrl and But mice. (B) Frequency and quantification of cell types in cytospins of BALF from control and butyrate-treated mice 18 hours after PAO1 infection; (C) Quantification of P. aeruginosa CFU in BALF 18 hours post-challenge. (D, E) Cytokine and chemokine production in BALF 18 hours following infection. (F) MFI of CXCR2 on lung AM and IM from Ctrl and But mice. (G) MFI of CXCR2 on Ctrl and But lung macrophages US or stimulated for 2 hours with LPS in vitro . (H) transmigration of Ctrl and But neutrophils toward a CXCL2 or CCL2 chemokine gradient after 3 hours. Results are a mean of two independent experiments. Values are expressed as mean ± standard error of mean; n = 6–14. Statistical significance was determined with one-way analysis of variance in (A–E, G, H) and Student’s t test (unpaired, two-tailed) in (F). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. AM = alveolar macrophages; BALF = bronchoalveolar lavage fluid; But = butyrate-treated; CFU = colony-forming units; Ctrl = control; IFN = interferon; IL = interleukin; IM = interstitial macrophages; LPS = lipopolysaccharide; MFI = mean fluorescence intensity; PAO1 = P. aeruginosa strain 1; TNF = tumor necrosis factor; US = unstimulated.

Article Snippet: To differentiate progenitor and neutrophil populations in the BM, cells were stained with antibodies to lineage cocktail, CD115 (BD Biosciences, clone 2B8), SiglecF (BD Biosciences, clone 2B8), CD117 BUV117 (BD Biosciences, clone 2B8), Sca1 APCFire (Biolegend, clone D7), FcgR biot (Miltenyi, clone 93), CD34 BV421 (Biolegend, clone SA376A4), CD11b BV510 (BD Biosciences, clone M1/70), Ly6G Pecy5 (eBioscience, clone 1A8-Ly6g), CXCR2 Pecy7 (Miltenyi, clone REA942) and streptavidin BUV395 (BD Biosciences).

Techniques: Control, Infection, In Vitro, Transmigration Assay, Two Tailed Test, Fluorescence

Journal: Advanced Science

Article Title: A Novel Self‐Regulated, Non‐Directional Magnetic Thermos‐Brachytherapy 125 I Seed Enhances Anticancer Efficacy by Rescuing Immune Escape

doi: 10.1002/advs.202508091

Figure Lengend Snippet: 125 I brachytherapy‐induced Cd274 + neutrophil recruitment promotes immune escape. A) Bar plot of the GSEA results showing functional differences in malignant epithelial cells between the radiotherapy/brachytherapy (R) and control (C) groups, with the bar length representing the normalized enrichment score. B) Dot plot showing differential expression levels and proportions of neutrophil chemokine‐related genes between the R and C groups. C) Violin plot of the differential expression of Cxcl2 between the R and C groups (Kruskal‐Wallis test). D) Violin plot showing the differential expression of Cxcr2 across general cell types. E) Stacked bar chart showing the proportions of neutrophil subtypes across different treatment groups. F) Dot plot illustrating the expression levels and proportions of functional markers in neutrophil subtypes. G) Flow cytometry analysis of subcutaneous tumors comparing the proportion of Cd274+ neutrophils between the C and R groups. Data are presented as mean ± s.d. (unpaired two‐sided t‐test, n = 5 per group). H) Representative images of multiplex immunofluorescence (mIF) staining showing colocalization of Cxcl2‐Cxcr2 interactions between malignant epithelial cells and neutrophils in subcutaneous tumors in the R and C groups (5 mice per group). I) Representative images of mIF staining comparing differential infiltration levels of CD274 + neutrophils in subcutaneous tumors between the R and C groups (5 mice per group). J) GSEA showing functional differences in malignant epithelial cells between the 125 I@MH (RM) and R groups. * : P < 0.05, ** : P < 0.01, *** : P < 0.001, **** : P < 0.0001.

Article Snippet: The primary antibodies included Epcam ( Cat : GB11274, Company : CiteAB, 1:2,000), Cxcl2 (16325‐1‐AP, Proteintech, 1:200), Csf3r (orb1676847, Biorbyt, 1:2,000), Cxcr2 (CSB‐ PA009567 , CUSABio, 1:200), Cd274/Pd‐l1 (28076‐1‐AP, Proteintech, 1:200), Cd3 (17‐0031‐82, Thermo Fisher, 1:200), Cd8 (11‐0081‐82, Thermo Fisher, 1:200), Ifn‐γ (12‐7311‐82, Thermo Fisher, 1:200), Pd‐1 (66220‐1‐lg, Proteintech, 1:200), F4/80 (123116, 1:100, BioLegend), Ccr2 (MAB55382, R&D Systems, 1:200), Apoe (66830‐1‐lg, Proteintech, 1:200).

Techniques: Functional Assay, Control, Quantitative Proteomics, Expressing, Flow Cytometry, Multiplex Assay, Immunofluorescence, Staining

Journal: Advanced Science

Article Title: A Novel Self‐Regulated, Non‐Directional Magnetic Thermos‐Brachytherapy 125 I Seed Enhances Anticancer Efficacy by Rescuing Immune Escape

doi: 10.1002/advs.202508091

Figure Lengend Snippet: MH rescues immune escape induced by 125 I brachytherapy. A) Stacked bar chart showing the proportions of lymphocyte subtypes across different treatment groups. B) Flow cytometry analysis of subcutaneous tumors comparing the proportions of exhausted T cells (upper) and effector T cells (lower) among the C, R, and RM groups. Data are presented as mean ± s.d. (unpaired two‐sided t ‐test, n = 5 per group). C) Representative image of multiplex immunofluorescence (mIF) staining comparing differential infiltration levels of exhausted T cells (upper) and effector T cells (lower) in subcutaneous tumors between the RM and R groups (5 mice per group). D) Comparison of subcutaneous tumor sizes in mice following no treatment, 125 I treatment, 125 I + anti‐PD‐1 treatment, and 125 I + anti‐PD‐1 + anti‐Cxcr2 treatment (6 mice per group). E) Stacked bar chart showing the proportions of phagocyte subtypes across different treatment groups. F) Bar plot of the GSEA results showing functional differences between Apoe + macrophages and Ccr2 + macrophages, with the bar length representing the normalized enrichment score. G) Flow cytometry analysis of subcutaneous tumors comparing the proportions of Ccr2 + macrophages (upper) and Apoe + macrophages (lower) among the C, R, and RM groups. Data are presented as mean ± s.d. (unpaired two‐sided t ‐test, n = 5 per group). H) Representative image of mIF staining comparing the differential infiltration levels of Ccr2 + macrophages (upper) and Apoe + macrophages (lower) in subcutaneous tumors between the RM and R groups. Data are presented as mean ± s.d. (unpaired two‐sided t ‐test, n = 5 per group). * : P < 0.05, ** : P < 0.01, *** : P < 0.001, **** : P < 0.0001.

Article Snippet: The primary antibodies included Epcam ( Cat : GB11274, Company : CiteAB, 1:2,000), Cxcl2 (16325‐1‐AP, Proteintech, 1:200), Csf3r (orb1676847, Biorbyt, 1:2,000), Cxcr2 (CSB‐ PA009567 , CUSABio, 1:200), Cd274/Pd‐l1 (28076‐1‐AP, Proteintech, 1:200), Cd3 (17‐0031‐82, Thermo Fisher, 1:200), Cd8 (11‐0081‐82, Thermo Fisher, 1:200), Ifn‐γ (12‐7311‐82, Thermo Fisher, 1:200), Pd‐1 (66220‐1‐lg, Proteintech, 1:200), F4/80 (123116, 1:100, BioLegend), Ccr2 (MAB55382, R&D Systems, 1:200), Apoe (66830‐1‐lg, Proteintech, 1:200).

Techniques: Flow Cytometry, Multiplex Assay, Immunofluorescence, Staining, Comparison, Functional Assay

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: Spinal injection of TNF-α-activated astrocytes induces mechanical allodynia via Cx43-mediated CXCL1 release. (A) Intrathecal injection of TNF-α-activated astrocytes elicited persistent mechanical allodynia for >48 h. Note this allodynia is reduced by pretreatment of astrocytes with Cx43 small interfering RNA (1 µg/ml, 18 h). *P < 0.05, compared with TNF-α or TNF-α + non-targeting control small interfering RNA treated group; n = 6 mice/group. (B) ELISA analysis shows increased CXCL1 release in the CSF at 3 h after the intrathecal injection of TNF-α-activated astrocytes. *P < 0.05, compared with vehcile group; #P < 0.05, compared with non-activated astrocytes; n = 4 mice/group. (C) Intrathecal injection of a CXCL1 neutralizing antibody (4 µg) transiently and partially reversed mechanical allodynia, induced by TNF-α-treated astrocytes. *P < 0.05, compared with control IgG group; n = 6 mice/group. (D) Intrathecal injection of the CXCR2 antagonist SB225002 (20 µg = 57 nmol) transiently and partially reversed mechanical allodynia, induced by TNF-α-activated astrocytes. *P < 0.05, compared with vehicle (PBS); n = 5–6 mice/group. All data are mean ± SEM. The differences between groups were analysed by ANOVA followed by Newman–Keuls test.

Article Snippet: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

Techniques: Injection, Small Interfering RNA, Control, Enzyme-linked Immunosorbent Assay

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: CXCL1, upregulated in spinal cord astrocytes after nerve injury, enhances excitatory synaptic transmission in spinal cord neurons and maintains neuropathic pain via CXCR2. (A) Western blotting shows CXCL1 upregulation in the spinal cord dorsal horn 21 days after CCI. Right, quantification of Cx43 levels in the dorsal horn. The western blot results are presented as a fold of sham control. *P < 0.05, compared to sham control, Student’s t-test, n = 4 mice/group. (B) Intrathecal injection of SB 225002 (20 µg), 21 days after CCI, reduced CCI-induced mechanical allodynia in the late phase. *P < 0.05, compared with vehicle (saline), Student’s t-test, n = 6 mice/group. (C) Double immunostaining of CXCL1 and GFAP in the dorsal horn 21 days after CCI. Note CXCL1 is primarily colocalized with GFAP. Arrows indicate doubled-labelled cells. Scale bar = 50 µm. (D and E) CXCL1 superfusion (100 ng/ml) increases spontaneous EPSC frequency (revealed by patch clamp recordings) in lamina IIo neurons of spinal cord slices. (E) Spontaneous EPSC frequency. *P < 0.05, Student’s t-test, n = 5 neurons/group. (F and G) CCI (21 d) increases spontaneous EPSC frequency, which is reversed by the CXCR2 antagonist SB225002 (1 µM). *P < 0.05, Student’s t-test, n = 5 neurons/group.

Article Snippet: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

Techniques: Transmission Assay, Western Blot, Control, Injection, Saline, Double Immunostaining, Patch Clamp

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: Schematic of working hypothesis for astrocytic Cx43-mediated late-phase neuropathic pain. CCI induces a persistent upregulation of Cx43 in spinal cord astrocytes. Cx43 expression and activity is also upregulated by TNF-α, secreted from microglia. Upregulation of Cx43 hemichannel activities results in CXCL1 release. Astrocytic CXCL1 secretion activates CXCR2 on neurons (central terminals of primary sensory neurons and spinal cord neurons), leading to enhanced excitatory synaptic transmission in nociceptive neurons (e.g. lamina IIo excitatory interneurons) and sustained neuropathic pain in the late-phase. Additionally, CXCL1 can also be secreted from intact or injured primary afferents in the spinal cord especially in the early phase of CCI.

Article Snippet: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

Techniques: Expressing, Activity Assay, Transmission Assay

Journal: Cell reports

Article Title: PAI-1-Dependent Inactivation of SMAD4-Modulated Junction and Adhesion Complex in Obese Endometrial Cancer

doi: 10.1016/j.celrep.2020.108253

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-CD182 conjugated to 147Sm (clone 5E8/CXCR2) , Fluidigm , Cat# 3147010B.

Techniques: Ubiquitin Proteomics, Recombinant, Multiplex Assay, Reverse Transcription, SYBR Green Assay, In Situ, shRNA, Software

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: a , Western blot analysis of CXCR2 expression in the negative control (NC), positive control (PC) and CM. Top: the NC is mouse embryonic fibroblasts, the PC is a human embryonic kidney cell line transfected with a Cxcr2 expression vector and the CM are cardiomyocytes of neonatal rats. Bottom: the NC is undifferentiated P19CL6 (mouse embryonic carcinoma) cells, the PC is a human embryonic kidney cell line transfected with a Cxcr2 expression vector and the CM are cardiomyocytes of mice. The experiment was repeated independently three times with similar results. b , Relative mRNA expression of HF markers in cardiomyocytes after addition of the medium conditioned by MYC overexpression in CFs, with and without a neutralizing antibody against CXCR2 ( n = 3 each). Data are shown as mean ± s.d. * P < 0.05; P < 0.001 (GFP ( n = 6) versus MYC ( n = 10), Nppa ), P = 0.0027 (MYC versus MYC plus anti-CXCR2 ( n = 5), Nppa ), P < 0.001 (GFP ( n = 7) versus MYC ( n = 10), Nppb ), P = 0.0027 (MYC versus MYC plus anti-CXCR2 ( n = 5), Nppb ), P < 0.001 (GFP ( n = 12) versus MYC ( n = 9), Myh7 ), P = 0.0040 (MYC versus MYC plus anti-CXCR2 ( n = 5), Myh7 ); significance was determined using the two-sided Mann–Whitney test for mean comparisons. c , Relative mRNA expression of HF markers measured by RT-qPCR in the hearts of control, TAC and TAC with neutralizing antibody against CXCR2. Data are shown as mean ± s.d. * P < 0.05; P < 0.001 (control ( n = 10) versus TAC ( n = 7), Nppa ), P = 0.017 (TAC versus TAC plus anti-CXCR2 ( n = 3), Nppa ), P = 0.0020 (control ( n = 10) versus TAC ( n = 4), Nppb ), P = 0.057 (TAC versus TAC plus anti-CXCR2 ( n = 3), Nppb ), P < 0.001 (control ( n = 10) versus TAC ( n = 6), Myh7 ), P = 0.038 (TAC versus TAC plus anti-CXCR2 ( n = 4), Myh7 ); significance was determined using the two-sided Mann–Whitney test for mean comparisons. d , Bar plots showing cardiac function evaluated by echocardiography in TAC mice with (anti-CXCR2) and without neutralizing antibody against CXCR2 (control). Data are shown as mean ± s.d. (TAC0w, n = 8 (control) and 7 (anti-CXCR2); TAC2w, 6 and 6; TAC4w, 6 and 6; TAC8w, 6 and 7; TAC12w, 8 and 7; TAC14w, 5 and 6; TAC16w, 7 and 6). * P < 0.05; P = 0.036 (TAC12w), P = 0.0037 (TAC14w), P = 0.015 (TAC16w); significance was determined using the two-sided Holm–Sidak test for mean comparisons. e , Comparison of cardiomyocyte size between control and TAC mice with and without neutralizing antibody against CXCR2 (anti-CXCR2). Cell size was evaluated using WGA staining. Quantitative analyses of the cell size in each mouse are shown ( n = 100 cells each). Box plots show the median (center line) and first and third quartiles (box edges), while the whiskers go from each quartile to the minimum or maximum. * P < 0.05; P < 0.001 (control versus TAC), P < 0.001 (TAC versus TAC + anti-CXCR2); significance was determined using the two-sided Mann–Whitney test for mean comparisons. f , Comparison of fibrotic areas between control and TAC mice with and without neutralizing antibody against CXCR2. Fibrosis was evaluated in the perivascular and interstitial regions by azan staining. Scale bars, 300 μm. Representative histological data and quantitative analyses of the fibrotic areas in each mouse are shown ( n = 7 each). Data are shown as mean ± s.d. * P < 0.05; P < 0.001 (control versus TAC, perivascular), P = 0.54 (TAC versus TAC plus anti-CXCR2, perivascular), P < 0.001 (control versus TAC, interstitial), P = 0.38 (TAC versus TAC plus anti-CXCR2, interstitial); significance was determined using the two-sided Mann–Whitney test for mean comparisons. g , Bar plots showing cardiac function evaluated by echocardiography in control, and shRNA of CXCR2-injected mice (shCXCR2) after TAC. Data are shown as mean ± s.d. ( n = 7 (control) and 5 (shCXCR2)). * P < 0.05; P = 0.025 (TAC8w); significance was determined using the two-sided Holm–Sidak test for mean comparisons. h , t -SNE plot of single-cell transcriptomes of human hearts (control: n = 1,673; HF: n = 29,496) (colored by cell clusters). Endo, endothelial cell; SM, smooth muscle cell; MC, myeloid cell; LC, lymphoid cell. i , t -SNE unsupervised clustering of fibroblasts in the human heart colored by cell clusters. j , Cell ratio of MYC -expressing CFs (control: n = 2; dilated cardiomyopathy (DCM): n = 4). Box plots show the median (center line) and first and third quartiles (box edges), while the whiskers go from each quartile to the minimum or maximum. k , Immunofluorescence for PDGFRα and smFISH for MYC and CXCL1 in human control hearts and DCM hearts. Scale bars, 20 μm. The experiment was repeated independently three times with similar results. The arrows indicate the colocalization of the MYC, CXCL1 and PDGFRα in the same cells. l , Bar plots showing the effect of CXCL1 on the contractile properties of iPSCM organoids. The plots show the rate of change of tissue contraction kinetics. Each parameter was normalized by the value of the control well ( n = 14 each). Data are shown as mean ± s.d. * P < 0.05; P = 0.047; significance was determined using the two-sided unpaired t -test for mean comparisons.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Western Blot, Expressing, Negative Control, Positive Control, Transfection, Plasmid Preparation, Over Expression, MANN-WHITNEY, Quantitative RT-PCR, Control, Comparison, Staining, shRNA, Injection, Immunofluorescence

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: Related to Fig. . a : Experimental scheme of TAC mice injected with a neutralising antibody against CXCR2. b : Bar plots showing left ventricle chamber size and wall thickness measured by echocardiography in control and anti-CXCR2 mice after TAC. Mean and standard error are shown (TAC0w, n = 8 [control] and 7 [anti-CXCR2]; TAC2w, 6 and 6; TAC4w, 6 and 6; TAC8w, 6 and 7; TAC12w, 8 and 7; TAC14w, 5 and 6; TAC16w, 7 and 6). *, p < 0.05; p = 0.040 (LVDs, TAC16w); significance was determined using the two-sided Holm-Sidak test for mean comparisons. Source data are provided as a Source Data file. c : Bar plots showing left ventricle chamber size and wall thickness measured by echocardiography in control and shCXCR2 mice after TAC. Mean and standard error are shown (n = 6 [control] and 5 [shCXCR2] for TAC0w, TAC2w, TAC4w, TAC6w, TAC8w, and TAC10w). *, p < 0.05; p = 0.015 (LVDd, TAC10w), p = 0.0020 (LVDs, TAC10w); significance was determined using the two-sided Holm-Sidak test for mean comparisons. Source data are provided as a Source Data file. d : Bar plots showing cardiac function, left ventricle chamber size, and wall thickness measured by echocardiography in control and shRNA of Cxcx1 injected mice (shCXCL1) after TAC. Mean and standard error are shown (n = 7 [control] and 5 [shCXCL1] for TAC0w, TAC2w, TAC4w, TAC6w, TAC8w, and TAC10w). *, p < 0.05; p = 0.0064 (LVDd, TAC10w); significance was determined using the two-sided Holm-Sidak test for mean comparisons. Source data are provided as a Source Data file. e : Left panel, Western blot analysis of p-ERK, ERK, and GAPDH expression in cardiomyocytes from neonatal rats treated with CXCL1 alone or in combination with the anti-CXCR2 antibody. Representative data are shown. Right panel, quantification of Western blot optical densities, with p-ERK/ERK ratios displayed. *, p < 0.05; significance was determined using the two-sided Mann-Whitney test for mean comparisons. Source data are provided as a Source Data file.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Injection, Control, shRNA, Western Blot, Expressing, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: Related to Fig. . a : Immunofluorescence for PDGFRα and single-molecule fluorescence in situ hybridization for MYC and CXCL1 in human dilated phase HCM (dHCM) and sarcoidosis hearts. Scale bars, 20 μm. Arrows indicate the colocalization of the MYC, CXCL1, and PDGFRα in the same cells. b : CXCR2 expression data using single-cell RNA-seq from our previous report. Horizontal axis, TNNT2 expression; vertical axis, CXCR2 expression. Black squares, cardiomyocytes from human control hearts; blue triangles, cardiomyocytes from human hearts of dilated cardiomyopathy. c : Western blot analysis of CXCR2 and GAPDH expression in iPSCM. d : Relative mRNA expression of HF markers in iPSCM (n = 3–5 each). Data are shown as mean ± SD. *, p < 0.05; p = 0.029 (Nppa), p = 0.036 (Nppb); significance was determined using the two-sided Mann-Whitney test for mean comparisons. Source data are provided as a Source Data file.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Immunofluorescence, Fluorescence, In Situ Hybridization, Expressing, RNA Sequencing, Control, Western Blot, MANN-WHITNEY

Journal: Nature Cardiovascular Research

Article Title: Heart failure-specific cardiac fibroblasts contribute to cardiac dysfunction via the MYC–CXCL1–CXCR2 axis

doi: 10.1038/s44161-025-00698-y

Figure Lengend Snippet: FACS analyses of CD3, CD11b, and Ly6G expression in mice hearts of sham (TAC0w), TAC4w, TAC8w, TAC12w, and TAC 12w with neutralising antibody against CXCR2.

Article Snippet: Nitrocellulose membranes with proteins were washed with 0.05% Tween-20 in Tris-buffered saline (TBST), blocked (ImmunoBlock; CTKN001, KAC) for 30 min at room temperature (20–25 °C) and then incubated with primary antibody (goat polyclonal anti-GAPDH antibody (1:1,000; sc20357, Santa Cruz Biotechnology), rabbit monoclonal anti-MYC antibody (1:1,000; ab32072, Abcam), anti-CXCR2 antibody (1:1,000; PA1029; Boster Biological Technology), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (1:1,000; 9101S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (137F5) antibody (1:1,000; 4695, Cell Signaling Technology) and CXCR2/CD182 Polyclonal Antibody (1:1000; Bioss)) at 4 °C overnight.

Techniques: Expressing