recombinant murine interferon gamma  (R&D Systems)

Journal: Advanced Science

Article Title: Pyruvate Kinase M2‐Responsive Release of Paclitaxel and Indoleamine 2,3‐Dioxygenase Inhibitor for Immuno‐Chemotherapy of Nonsmall Cell Lung Cancer

doi: 10.1002/advs.202409790

Figure Lengend Snippet: Immune activation by GP@ Gel Nap‐T in tumor tissues in vivo. a) Schematic illustration about the workflow of FCM and IF analysis of tumor tissues from CMT167 tumor‐bearing mice on day 14 after different treatments. b–d) Quantitative FCM analysis of the proportions of (b) CD4 + T cells, (c) CD8 + T cells, (d) NK cells in the tumor tissues from CMT167 tumor‐bearing mice on day 14 after different treatments ( n = 5 biological independent samples). e) Representative IF staining of CD4, CD8, and NK1.1 in the tumor tissues from CMT167 tumor‐bearing mice on day 14 after different treatments. DAPI (blue), CD4 (green), CD8, and NK1.1 (red). Scale bars: 50 µm ( n = 3 biological independent samples). f–q) Quantitative FCM analysis of the proportions of (f) M1‐TAMs, (g) M2‐TAMs, (h) M‐MDSCs, (i) PMN‐MDSCs, (j) Tregs, (k) Ki‐67 + CD8 + T cells, (l) GZB + CD8 + T cells, (m) GZB + NK1.1 + cells, (n) IFN‐γ + CD8 + T cells, (o) IFN‐γ + NK1.1 + cells, (p) CD8 + T CM (CD8 + CD44 + CD62L + ) cells, and (q) CD8 + T EM (CD8 + CD44 + CD62L − ) in the tumor tissues from CMT167 tumor‐bearing mice on day 14 after different treatments ( n = 5 biological independent samples). Results are presented as mean ± SD. Statistical significance was assessed using one‐way ANOVA with Tukey's post‐test. ns: no significant difference, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Assay kits for methyl thiazolyl tetrazolium (MTT) cytotoxicity, glutathione (GSH), adenosine 5'‐triphosphate (ATP), one‐step TUNEL apoptosis assay kit, anti‐mouse calreticulin (CRT) and anti‐mouse Ki‐67 primary antibodies, DNase I, recombinant murine interleukin‐4 (IL‐4), recombinant murine interferon‐gamma (IFN‐γ), RIPA lysis buffer, PI, and phenylmethylsulfonyl fluoride (PMSF) were all obtained from Beyotime (Shanghai, China).

Techniques: Activation Assay, In Vivo, Staining

Journal: Advanced Science

Article Title: Pyruvate Kinase M2‐Responsive Release of Paclitaxel and Indoleamine 2,3‐Dioxygenase Inhibitor for Immuno‐Chemotherapy of Nonsmall Cell Lung Cancer

doi: 10.1002/advs.202409790

Figure Lengend Snippet: Immune activation by GP@Gel Nap‐T in TDLNs in vivo. a) Schematic illustration about the workflow of FCM and IF analysis of TDLNs from CMT167 tumor‐bearing mice on day 14 after different treatments. b–d) Quantitative FCM analysis of the proportions of (b) CD4 + T cells, (c) CD8 + T cells, (d) NK cells in TDLNs from CMT167 tumor‐bearing mice on day 14 after different treatments ( n = 5 biological independent samples). e) Representative IF staining of CD4, CD8, and NK1.1 in TDLNs from CMT167 tumor‐bearing mice on day 14 after different treatments. DAPI (blue), CD4 (green), CD8, and NK1.1 (red). Scale bars: 50 µm ( n = 3 biological independent samples). f–q) Quantitative FCM analysis of the proportions of (f) M1‐TAMs, (g) M2‐TAMs, (h) M‐MDSCs, (i) PMN‐MDSCs, (j) Tregs, (k) Ki‐67 + CD8 + T cells, (l) GZB + CD8 + T cells, (m) IFN‐γ + CD8 + T cells, (n) GZB + NK1.1 + cells, (o) IFN‐γ + NK1.1 + cells, (p) CD8 + T CM (CD8 + CD44 + CD62L + ) cells, and (q) CD8 + T EM (CD8 + CD44 + CD62L − ) in TDLNs from CMT167 tumor‐bearing mice on day 14 after different treatments ( n = 5 biological independent samples). Results are presented as mean ± SD. Statistical significance was assessed using one‐way ANOVA with Tukey's post‐test. ns: no significant difference, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Assay kits for methyl thiazolyl tetrazolium (MTT) cytotoxicity, glutathione (GSH), adenosine 5'‐triphosphate (ATP), one‐step TUNEL apoptosis assay kit, anti‐mouse calreticulin (CRT) and anti‐mouse Ki‐67 primary antibodies, DNase I, recombinant murine interleukin‐4 (IL‐4), recombinant murine interferon‐gamma (IFN‐γ), RIPA lysis buffer, PI, and phenylmethylsulfonyl fluoride (PMSF) were all obtained from Beyotime (Shanghai, China).

Techniques: Activation Assay, In Vivo, Staining