cxcl12 staining  (R&D Systems)

Journal: Redox Biology

Article Title: Hydrogen sulfide inhibits recruitment of monocyte-derived tumor associated macrophages in glioblastoma by downregulating CXCL12

doi: 10.1016/j.redox.2025.103866

Figure Lengend Snippet: Histologic sections of IDH wildtype glioblastoma with low H 2 S concentrations (A, D, and G) and high H 2 S concentrations (B, E, and H) were immunostained with antibody against a TAM marker CD163 (A and B), combination of antibodies against a pan-myeloid marker Iba1 (green, D and E) and resident microglia specific marker P2RY12 (red, D and E), and antibody against chemokine CXCL12 (G and H). There was an inverse correlation between tumor H 2 S concentrations and density of CD163-positive TAMs (C; n = 10, ∗P = 0.044, Spearman), percent of Iba1-positive, P2RY12-negative, peripherally derived TAMs (F; n = 10, ∗P = 0.043, Spearman), and density of CXCL12 expressing cells (I; n = 10, ∗P = 0.0005, Spearman). Scale bar, 50 μm.

Article Snippet: Following heat-induced citrate buffer antigen retrieval and peroxide treatment, paraffin sections of human or mice tumors were incubated at 4 °C overnight with the following antibodies: rabbit anti-human CD163 (1:200, ab87099, Abcam), rabbit anti-human/mouse CXCL12 (1:50, MAB350, R&D Systems), rabbit anti-mouse ionized calcium binding adaptor molecule 1 (Iba1; 1:500, NBP2-19019, Novus Biologicals), or rabbit anti-human hypoxia-inducible factor-1 (HIF-1; 1:100, ab51608, Abcam).

Techniques: Marker, Derivative Assay, Expressing

Journal: Redox Biology

Article Title: Hydrogen sulfide inhibits recruitment of monocyte-derived tumor associated macrophages in glioblastoma by downregulating CXCL12

doi: 10.1016/j.redox.2025.103866

Figure Lengend Snippet: (A) Wildtype mice fed a diet containing H 2 S donor SG1002 (40 mg/kg/d; n = 5) demonstrated increased plasma free H 2 S concentrations compared to those fed a control diet (n = 5; ∗∗∗∗P < 0.0001, t -test). (B) Similarly, relative plasma SSP4 fluorescence (indicating sulfane sulfur concentrations) was increased in the SG1002 group (n = 4) compared to controls (n = 4, ∗P = 0.016, t -test). (C) Kaplan-Meier curves showing overall survival for Nestin-TVA mice injected with RCAS-PDGFB and RCAS-STAT3 and fed a control (n = 25) or SG1002-containing diet (n = 25, ∗P = 0.020, log-rank test). Histologic sections of tumors from control (D, G, and J) and SG1002 groups (E, H, and K) were immunostained with an antibody against Iba1 (D and E), combination of antibodies against a pan-myeloid marker Iba1 (green, G and H) and resident microglia specific marker P2RY12 (red, G and H), and antibody against chemokine CXCL12 (J and K). SG1002 administration decreased accumulation of Iba1-positive TAMs (F, n = 4, ∗P = 0.013, t -test), relative abundance of Iba1-positive, P2RY12-negative, peripherally derived TAMs (I: n = 4, ∗∗P = 0.0018, t -test) and density of CXCL12 expressing cells (L, n = 3–4, ∗P = 0.036, t -test). Scale bar, 50 μm.

Article Snippet: Following heat-induced citrate buffer antigen retrieval and peroxide treatment, paraffin sections of human or mice tumors were incubated at 4 °C overnight with the following antibodies: rabbit anti-human CD163 (1:200, ab87099, Abcam), rabbit anti-human/mouse CXCL12 (1:50, MAB350, R&D Systems), rabbit anti-mouse ionized calcium binding adaptor molecule 1 (Iba1; 1:500, NBP2-19019, Novus Biologicals), or rabbit anti-human hypoxia-inducible factor-1 (HIF-1; 1:100, ab51608, Abcam).

Techniques: Clinical Proteomics, Control, Fluorescence, Injection, Marker, Derivative Assay, Expressing

Journal: Redox Biology

Article Title: Hydrogen sulfide inhibits recruitment of monocyte-derived tumor associated macrophages in glioblastoma by downregulating CXCL12

doi: 10.1016/j.redox.2025.103866

Figure Lengend Snippet: (A) Transwell migration of THP-1 macrophages in response to conditioned media (CM) from T98G glioblastoma cells treated with PBS or 100 μM Na 2 S 4 , as well as serum-free media (NC) or media containing 10 % serum (PC). (B) CM from PBS treated T98G cells promoted chemotaxis of THP-1 macrophages compared to NC (∗∗P = 0.0070, ANOVA with Tukey's post hoc test), but this effect was abrogated with CM from Na 2 S 4 treated T98G cells. (∗P = 0.010, ANOVA with Tukey's post hoc test). Data are representative of 6 independent experiments per each condition. Scale bar, 50 μm. Na 2 S 4 treatment of T98G cells decreased CXCL12 transcript levels (C) and CXCL12 concentrations in CM (D) compared to PBS treatment (n = 3, ∗P = 0.010, ∗∗∗∗P < 0.0001, t -test).

Article Snippet: Following heat-induced citrate buffer antigen retrieval and peroxide treatment, paraffin sections of human or mice tumors were incubated at 4 °C overnight with the following antibodies: rabbit anti-human CD163 (1:200, ab87099, Abcam), rabbit anti-human/mouse CXCL12 (1:50, MAB350, R&D Systems), rabbit anti-mouse ionized calcium binding adaptor molecule 1 (Iba1; 1:500, NBP2-19019, Novus Biologicals), or rabbit anti-human hypoxia-inducible factor-1 (HIF-1; 1:100, ab51608, Abcam).

Techniques: Migration, Chemotaxis Assay