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pex3 overexpression lentiviruses  (Genechem)
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Genechem pex3 overexpression lentiviruses
CCL7 activates <t>PEX3</t> to regulate fatty acid metabolism. ( A-C ) Volcano plot ( A ) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis ( B ) of differentially expressed proteins in Ccl7 f/f and Ccl7 MKO TAMs derived from MC38 tumors, along with a heatmap of peroxisome-related differentially expressed proteins ( C ), with n = 3. ( D ) The expression levels of peroxisome-related genes in TAMs from Ccl7 f/f and Ccl7 MKO tumor tissues (n = 3). ( E ) The expression levels of Pex3 in BMDMs from Ccl7 f/f and Ccl7 MKO mice were assessed using western blot analysis. ( F ) Representative images of PMP70 staining (red) in TAMs from Ccl7 f/f and Ccl7 MKO tumor tissues, with a scale bar of 5 µm. ( G ) PMP70 expression in BMDMs from both Ccl7 f/f and Ccl7 MKO mice, as well as BMDMs stimulated with recombinant Ccl7 for 24 hours. ( H ) BMDMs were transfected with siRNA targeting Pex3 , and OCR and SRC were measured (n = 5). ( I ) BMDMs transfected with si Pex3 and subsequently stimulated with Ccl7 for 24 hours exhibited ROS production, which was determined by FCM (n = 4). ( J ) iNOS and Arg1 expression in BMDMs transfected with siPex3 was assessed by FCM (n = 4). ( K, L ) CD8 + T lymphocytes were co-cultured with either siNC or si Pex3 TAMs at a 2:1 ratio; the proliferation of CD8 + T cells induced by anti-CD3 and anti-CD28 was measured by FCM ( K ), and the production of IFN-γ in CD8 + T cells was measured by FCM (L; n = 4). ( M ) Intracellular staining of p-Akt1 in TAMs from Ccl7 f/f or Ccl7 MKO tumor-bearing mice (n=5). ( N ) BMDMs were pretreated with or without BEZ235 (200 nM). and subsequently stimulated with Ccl7 for 24 hours, Arg1 expression was determined by FCM, n=4. ( O, P ) BMDMs were treated with or without BEZ235 (200 nM), and the expression of Pex3 and Acox1 was quantified using western blot analysis ( O ). ( P ) PMP70 expression in BMDMs was assessed ( P ). ( Q, R ) BMDMs were transfected with siRNA targeting Ccr1, Ccr2, and Ccr3, and the expression of Arg1 ( Q ) and iNOS ( R ) was analyzed using FCM. Data are presented as mean±SD. Statistical significance was determined using the Mann-Whitney test, with *p<0.05, **p<0.01, ***p<0.001, and ns indicating no significant difference. BMDM, bone marrow-derived macrophages; FCM, flow cytometry; IFN, interferon; TAMs, tumor-associated macrophages; KEGG, Kyoto Encyclopedia of Genes and Genomes; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
Pex3 Overexpression Lentiviruses, supplied by Genechem, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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CCL7 activates PEX3 to regulate fatty acid metabolism. ( A-C ) Volcano plot ( A ) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis ( B ) of differentially expressed proteins in Ccl7 f/f and Ccl7 MKO TAMs derived from MC38 tumors, along with a heatmap of peroxisome-related differentially expressed proteins ( C ), with n = 3. ( D ) The expression levels of peroxisome-related genes in TAMs from Ccl7 f/f and Ccl7 MKO tumor tissues (n = 3). ( E ) The expression levels of Pex3 in BMDMs from Ccl7 f/f and Ccl7 MKO mice were assessed using western blot analysis. ( F ) Representative images of PMP70 staining (red) in TAMs from Ccl7 f/f and Ccl7 MKO tumor tissues, with a scale bar of 5 µm. ( G ) PMP70 expression in BMDMs from both Ccl7 f/f and Ccl7 MKO mice, as well as BMDMs stimulated with recombinant Ccl7 for 24 hours. ( H ) BMDMs were transfected with siRNA targeting Pex3 , and OCR and SRC were measured (n = 5). ( I ) BMDMs transfected with si Pex3 and subsequently stimulated with Ccl7 for 24 hours exhibited ROS production, which was determined by FCM (n = 4). ( J ) iNOS and Arg1 expression in BMDMs transfected with siPex3 was assessed by FCM (n = 4). ( K, L ) CD8 + T lymphocytes were co-cultured with either siNC or si Pex3 TAMs at a 2:1 ratio; the proliferation of CD8 + T cells induced by anti-CD3 and anti-CD28 was measured by FCM ( K ), and the production of IFN-γ in CD8 + T cells was measured by FCM (L; n = 4). ( M ) Intracellular staining of p-Akt1 in TAMs from Ccl7 f/f or Ccl7 MKO tumor-bearing mice (n=5). ( N ) BMDMs were pretreated with or without BEZ235 (200 nM). and subsequently stimulated with Ccl7 for 24 hours, Arg1 expression was determined by FCM, n=4. ( O, P ) BMDMs were treated with or without BEZ235 (200 nM), and the expression of Pex3 and Acox1 was quantified using western blot analysis ( O ). ( P ) PMP70 expression in BMDMs was assessed ( P ). ( Q, R ) BMDMs were transfected with siRNA targeting Ccr1, Ccr2, and Ccr3, and the expression of Arg1 ( Q ) and iNOS ( R ) was analyzed using FCM. Data are presented as mean±SD. Statistical significance was determined using the Mann-Whitney test, with *p<0.05, **p<0.01, ***p<0.001, and ns indicating no significant difference. BMDM, bone marrow-derived macrophages; FCM, flow cytometry; IFN, interferon; TAMs, tumor-associated macrophages; KEGG, Kyoto Encyclopedia of Genes and Genomes; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Journal: Journal for Immunotherapy of Cancer

Article Title: Macrophage CCL7 promotes resistance to immunotherapy for colorectal cancer by regulating the infiltration of macrophages and CD8+ T cells

doi: 10.1136/jitc-2025-013027

Figure Lengend Snippet: CCL7 activates PEX3 to regulate fatty acid metabolism. ( A-C ) Volcano plot ( A ) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis ( B ) of differentially expressed proteins in Ccl7 f/f and Ccl7 MKO TAMs derived from MC38 tumors, along with a heatmap of peroxisome-related differentially expressed proteins ( C ), with n = 3. ( D ) The expression levels of peroxisome-related genes in TAMs from Ccl7 f/f and Ccl7 MKO tumor tissues (n = 3). ( E ) The expression levels of Pex3 in BMDMs from Ccl7 f/f and Ccl7 MKO mice were assessed using western blot analysis. ( F ) Representative images of PMP70 staining (red) in TAMs from Ccl7 f/f and Ccl7 MKO tumor tissues, with a scale bar of 5 µm. ( G ) PMP70 expression in BMDMs from both Ccl7 f/f and Ccl7 MKO mice, as well as BMDMs stimulated with recombinant Ccl7 for 24 hours. ( H ) BMDMs were transfected with siRNA targeting Pex3 , and OCR and SRC were measured (n = 5). ( I ) BMDMs transfected with si Pex3 and subsequently stimulated with Ccl7 for 24 hours exhibited ROS production, which was determined by FCM (n = 4). ( J ) iNOS and Arg1 expression in BMDMs transfected with siPex3 was assessed by FCM (n = 4). ( K, L ) CD8 + T lymphocytes were co-cultured with either siNC or si Pex3 TAMs at a 2:1 ratio; the proliferation of CD8 + T cells induced by anti-CD3 and anti-CD28 was measured by FCM ( K ), and the production of IFN-γ in CD8 + T cells was measured by FCM (L; n = 4). ( M ) Intracellular staining of p-Akt1 in TAMs from Ccl7 f/f or Ccl7 MKO tumor-bearing mice (n=5). ( N ) BMDMs were pretreated with or without BEZ235 (200 nM). and subsequently stimulated with Ccl7 for 24 hours, Arg1 expression was determined by FCM, n=4. ( O, P ) BMDMs were treated with or without BEZ235 (200 nM), and the expression of Pex3 and Acox1 was quantified using western blot analysis ( O ). ( P ) PMP70 expression in BMDMs was assessed ( P ). ( Q, R ) BMDMs were transfected with siRNA targeting Ccr1, Ccr2, and Ccr3, and the expression of Arg1 ( Q ) and iNOS ( R ) was analyzed using FCM. Data are presented as mean±SD. Statistical significance was determined using the Mann-Whitney test, with *p<0.05, **p<0.01, ***p<0.001, and ns indicating no significant difference. BMDM, bone marrow-derived macrophages; FCM, flow cytometry; IFN, interferon; TAMs, tumor-associated macrophages; KEGG, Kyoto Encyclopedia of Genes and Genomes; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Pex3 overexpression lentiviruses were obtained from GeneChem (Shanghai, China).

Techniques: Derivative Assay, Expressing, Western Blot, Staining, Recombinant, Transfection, Cell Culture, MANN-WHITNEY, Flow Cytometry

Targeting CCL7 enhances the efficacy of immunotherapy with anti-PD-L1 antibody. ( A-G ) MC38-luc and CT26-luc cells were intraperitoneally injected into mice, which subsequently received 100 µg of anti-Ccl7 antibody (α-Ccl7) or Veh (PBS) every 3 days, starting on day 7 post-tumor injection. Tumor growth was monitored ( A ), and survival of tumor-bearing mice was assessed (B, n = 9–12). The percentage of TAMs was determined ( C ), along with the expression of iNOS and Arg1 in macrophages ( D ), the presence of MHC II + TAMs ( E ), Stat1 expression in TAMs ( F ), and the frequency of IFNγ + and TNF-α + CD8 + T cells in tumors ( G ), with n = 5 for these analyses. ( H-L ) C57 and BALB/c mice were intraperitoneally injected with MC38-luc and CT26-luc cells, respectively. Tumor-bearing mice were injected with Veh, α-Ccl7, anti-PD-L1 (αPD-L1), or a combination of α-Ccl7 and anti-PD-L1 (α-C+α-P). Tumor growth was monitored ( H ), survival of tumor-bearing mice was analyzed (I, n = 10–14), and the percentages of TAMs ( J ), CD8 + T cells ( K ), and IFNγ + CD8 + T cells ( L ) in the tumor tissues were evaluated using FCM (n = 5). ( M ) Correlation between CCL7 and CD163 , MRC1 , and PEX3 in human CRC tissues. Data are presented as mean±SD. Statistical significance was determined using the Mann-Whitney test ( A, C-G ), log-rank (Mantel-Cox) test ( B and I ), and one-way or two-way analysis of variance with Sidak multiple comparisons test ( H, J-L ), with significance levels indicated as *p<0.05, **p<0.01, ***p<0.001. CRC, colorectal cancer; IFN, interferon; TAMs, tumor-associated macrophages.

Journal: Journal for Immunotherapy of Cancer

Article Title: Macrophage CCL7 promotes resistance to immunotherapy for colorectal cancer by regulating the infiltration of macrophages and CD8+ T cells

doi: 10.1136/jitc-2025-013027

Figure Lengend Snippet: Targeting CCL7 enhances the efficacy of immunotherapy with anti-PD-L1 antibody. ( A-G ) MC38-luc and CT26-luc cells were intraperitoneally injected into mice, which subsequently received 100 µg of anti-Ccl7 antibody (α-Ccl7) or Veh (PBS) every 3 days, starting on day 7 post-tumor injection. Tumor growth was monitored ( A ), and survival of tumor-bearing mice was assessed (B, n = 9–12). The percentage of TAMs was determined ( C ), along with the expression of iNOS and Arg1 in macrophages ( D ), the presence of MHC II + TAMs ( E ), Stat1 expression in TAMs ( F ), and the frequency of IFNγ + and TNF-α + CD8 + T cells in tumors ( G ), with n = 5 for these analyses. ( H-L ) C57 and BALB/c mice were intraperitoneally injected with MC38-luc and CT26-luc cells, respectively. Tumor-bearing mice were injected with Veh, α-Ccl7, anti-PD-L1 (αPD-L1), or a combination of α-Ccl7 and anti-PD-L1 (α-C+α-P). Tumor growth was monitored ( H ), survival of tumor-bearing mice was analyzed (I, n = 10–14), and the percentages of TAMs ( J ), CD8 + T cells ( K ), and IFNγ + CD8 + T cells ( L ) in the tumor tissues were evaluated using FCM (n = 5). ( M ) Correlation between CCL7 and CD163 , MRC1 , and PEX3 in human CRC tissues. Data are presented as mean±SD. Statistical significance was determined using the Mann-Whitney test ( A, C-G ), log-rank (Mantel-Cox) test ( B and I ), and one-way or two-way analysis of variance with Sidak multiple comparisons test ( H, J-L ), with significance levels indicated as *p<0.05, **p<0.01, ***p<0.001. CRC, colorectal cancer; IFN, interferon; TAMs, tumor-associated macrophages.

Article Snippet: Pex3 overexpression lentiviruses were obtained from GeneChem (Shanghai, China).

Techniques: Injection, Expressing, MANN-WHITNEY