mouse anti mertk  (Santa Cruz Biotechnology)

Journal: International immunopharmacology

Article Title: Immune checkpoint inhibitor induces cardiac injury by impairing efferocytosis of macrophages via MerTK cleavage.

doi: 10.1016/j.intimp.2025.114263

Figure Lengend Snippet: Fig. 3. MerTK took effect in cardiac injury elicited by the PD-1 inhibitor. (A) Immunofluorescent double staining of the section of frozen mouse heart with antibodies against CD68(red) and MerTK (green), DAPI-stained cellular nuclei, Scale bar = 20 μm. (B) Quantitative analysis of the CD68 and MerTK expression. (C) Immu nohistochemistry image of MerTK staining in a section of frozen mouse heart (×10, Scale bar = 200 μm, ×40, Scale bar = 50 μm). (D) Quantitative analysis of the immunohistochemistry positive cells, n = 3 mice in each group. (E–F) Images and quantitative analysis of tissues protein MerTK, n = 3 mice in each group. (G–H) Images and quantitative analysis of MerTK and SolMer protein expression levels in RAW264.7 cells treated with PD-1 inhibitor at different times (0, 2, 4, 6, 12, and 24 h). (I) Immunofluorescent double staining of the RAW264.7 cells with antibodies against CD68(red) and MerTK(green), DAPI-stained cellular nuclei, Scale bar = 50 μm. (J) Quantitative analysis of the CD68 and MerTK expression. (K) Serum levels of SolMer in the mice, n = 12 mice in each group. (L) ELISA analysis of SolMer in medium of RAW264.7 cells treated with PD-1 inhibitor. (M) mRNA expression of MerTK in mouse cardiac tissue, n = 3 mice in each group. (N) mRNA expression of MerTK in RAW264.7 cells. Values in plots are expressed as means SD. *p < 0.05, **p < 0.01, ***p < 0.001and ****p < 0.0001, ns, not significant.

Article Snippet: The cells or sections were analyzed using immunofluorescence staining with primary antibodies, including rabbit CD68 (1:200, Abcam), mouse MerTK (1:50, Santa Cruz), rat F4/80 (1:200, Abcam), and rabbit ADAM17 (1:200, Abcam) at 4 ◦C overnight.

Techniques: Double Staining, Staining, Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: International immunopharmacology

Article Title: Immune checkpoint inhibitor induces cardiac injury by impairing efferocytosis of macrophages via MerTK cleavage.

doi: 10.1016/j.intimp.2025.114263

Figure Lengend Snippet: Fig. 4. PD-1 inhibitor induces MerTK cleavage through upregulating ADAM17. (A) Immunofluorescent double staining of the frozen heart tissue with antibodies against F4/80 (green) and ADAM17 (red), DAPI-stained cellular nuclei, Scale bar = 20 μm. (B) Quantitative analysis of the F4/80 and ADAM17 expression, n = 3 mice in each group. (C) Immunohistochemistry image of ADAM17 staining in frozen mouse heart (×10, Scale bar = 200 μm, ×40, Scale bar = 50 μm. (D) Quan titative analysis of the immunohistochemistry positive cells, n = 3 mice in each group. (E–F) Images and quantitative analysis of tissues protein ADAM17, n = 3 mice in each group. (G–I) Images and quantitative analysis of MerTK and ADAM17 protein expression levels in RAW264.7 cells. (J) Immunofluorescent double staining of the RAW264.7 cells with antibodies against F4/80(green) and ADAM17(red), DAPI-stained cellular nuclei, Scale bar = 50 μm. (K) Quantitative analysis of the CD68 and MerTK expression. Values in plots are expressed as means SD. *p < 0.05, **p < 0.01, ***p < 0.001and ****p < 0.0001, ns, not significant.

Article Snippet: The cells or sections were analyzed using immunofluorescence staining with primary antibodies, including rabbit CD68 (1:200, Abcam), mouse MerTK (1:50, Santa Cruz), rat F4/80 (1:200, Abcam), and rabbit ADAM17 (1:200, Abcam) at 4 ◦C overnight.

Techniques: Double Staining, Staining, Expressing, Immunohistochemistry

Journal: International immunopharmacology

Article Title: Immune checkpoint inhibitor induces cardiac injury by impairing efferocytosis of macrophages via MerTK cleavage.

doi: 10.1016/j.intimp.2025.114263

Figure Lengend Snippet: Fig. 5. The ADAM17 inhibitor TAPI-0 restored the ability of macrophages to phagocytose apoptotic cells. (A–B) Images and quantitative analysis of MerTK and SolMer protein expression levels in RAW264.7 cells pretreated with the ADAM17 inhibitor TAPI-0 (10 μM) for 2 h, and then intervened with PD-1 inhibitor for 6 h. (C) Fluorescent images for phagocytosis of apoptotic cells by macrophages, ACs: Apoptotic Cells, Scale bar = 50 μm, Scale bar in enlarge = 20 μm. (D) Efferocytosis evaluation based on the fluorescent image. (E) Flow cytometry evaluation for phagocytosis of apoptotic cells by macrophages, ACs, Apoptotic Cells. (F) Efferocytosis index based on flow fluorescence intensity. Values in plots are expressed as means SD. *p < 0.05, **p < 0.01, ***p < 0.001and ****p < 0.0001, ns, not significant.

Article Snippet: The cells or sections were analyzed using immunofluorescence staining with primary antibodies, including rabbit CD68 (1:200, Abcam), mouse MerTK (1:50, Santa Cruz), rat F4/80 (1:200, Abcam), and rabbit ADAM17 (1:200, Abcam) at 4 ◦C overnight.

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: International immunopharmacology

Article Title: Immune checkpoint inhibitor induces cardiac injury by impairing efferocytosis of macrophages via MerTK cleavage.

doi: 10.1016/j.intimp.2025.114263

Figure Lengend Snippet: Fig. 6. PD-1 inhibitor activates ADAM17 through phosphorylation of the MKK3/P38 MAPK pathway. (A–B) Images and quantitative analysis of tissues protein MKK3, p-MKK3, P38 and p-P38, n = 3 mice in each group. (C–D) Images and quantitative analysis of MKK3, p-MKK3, P38 and p-P38 protein expression levels in RAW264.7 cells. (E–F) Images and quantitative analysis of ADAM17 protein expression levels in RAW264.7 cells pretreated with P38 inhibitor SB203580 (20 μM) for 2 h, and then intervened with PD-1 inhibitor for 6 h. (G–H) Images and quantitative analysis of MerTK and SolMer protein expression levels in RAW264.7 cells pretreated with P38 inhibitor SB203580 (20 μM) for 2 h, and then intervened with PD-1 inhibitor for 6 h.2 h before PD-1 inhibitor exposure for 6 h. Values in plots are expressed as means SD. *p < 0.05, **p < 0.01, ***p < 0.001and ****p < 0.0001, ns, not significant.

Article Snippet: The cells or sections were analyzed using immunofluorescence staining with primary antibodies, including rabbit CD68 (1:200, Abcam), mouse MerTK (1:50, Santa Cruz), rat F4/80 (1:200, Abcam), and rabbit ADAM17 (1:200, Abcam) at 4 ◦C overnight.

Techniques: Phospho-proteomics, Expressing

Journal: International immunopharmacology

Article Title: Immune checkpoint inhibitor induces cardiac injury by impairing efferocytosis of macrophages via MerTK cleavage.

doi: 10.1016/j.intimp.2025.114263

Figure Lengend Snippet: Fig. 8. PD-1 inhibitor acts by binding to the PD-1 receptor on the surface of macrophages. (A–B) Images and quantitative analysis of PD-1 protein expression levels in RAW264.7 cells. (C) mRNA expression of PD-1 in RAW264.7 cells. (D–F) Images and quantitative analysis of MerTK and ADAM17 protein expression levels in RAW264.7 cells. (G–H) Images and quantitative analysis of MKK3 and p-MKK3 protein expression levels in RAW264.7 cells. (I–J) Images and quantitative analysis of P38 and p-P38 protein expression levels in RAW264.7 cells. (sh-NC: The RAW264.7 was transfected with the control lentivirus; sh-PD-1: The RAW264.7 was transfected with shRNA PD-1) Values in plots are expressed as means SD. *p < 0.05, **p < 0.01, ***p < 0.001and ****p < 0.0001, ns, not significant.

Article Snippet: The cells or sections were analyzed using immunofluorescence staining with primary antibodies, including rabbit CD68 (1:200, Abcam), mouse MerTK (1:50, Santa Cruz), rat F4/80 (1:200, Abcam), and rabbit ADAM17 (1:200, Abcam) at 4 ◦C overnight.

Techniques: Binding Assay, Expressing, Transfection, Control, shRNA