Journal: European Journal of Inflammation

Article Title: Correlation between circulating myeloid-derived suppressor cells and serum miRNA21 in non-small cell lung cancer and its clinical significance

doi: 10.1177/1721727x231175019

Figure Lengend Snippet: Figure 3. The effect of miRNA-21 on MDSCs. MDSC cells were transfected with control mimics, miR-21 mimics, or control inhibtor and miR-21 inhibtor. (a) miRNA-21 expressions of four groups were examined by q-PCR. (b) The transfected cell viability was assessed using MTT. (c) The level of ARG-1 was detected by ELISA assay. (d) The level of iNOS was detected by ELISA assay. Unpaired t-tests was used for comparisons between two groups. **p < 0.01, ***p < 0.001, miR-21 mimics group was significantly different from the values in the control mimics group. #p < 0.05, ##p < 0.01, ###p < 0.001, miR-21 inhibtor group was significantly different from the values in the control inhibtor group.

Article Snippet: The iNOS (CSBEL002005HU) and ARG-1 (CSB-E08148h) detection kits were purchased from Cusabio, Wuhan, China (refer to the kit manual for the specific protocol).

Techniques: Transfection, Control, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Breast Cancer Macrophage Heterogeneity and Self-renewal are Determined by Spatial Localization

doi: 10.1101/2023.10.24.563749

Figure Lengend Snippet: ( a ) Immunoblotting for the cleaved Notch intracellular domain (NICD), ARG1, phospho-PI3K (pAKT), and phospho-MAPK (pERK), confirming active Notch and PI3K pathways in oTME macrophages but not in CSF-1 macrophages. ( b ) Impact of γ-secretase inhibition on proliferation of oTME macrophages. BM monocytes were isolated and plated with oTME cells for 7 days and then were treated with either compound-E (CompE; 10μM) or DMSO as a control for additional 10 days. Macrophages and oTME cells were analyzed by flow cytometry for Ki67 expression. Data (n=3) are shown as mean±SD, Welch’s t- test. ( c ) EdU incorporation in BMDMs following treatment with Adam17 protease inhibitor (A17Pro) or PBS as vehicle control. Cells were plated with oTME cells in the presence of A17Pro or PBS and cultured for seven days. Cell cultures were then labeled with EdU for 48hrs and were analyzed by flow cytometry for EdU incorporation (n=4 replicates). Data are shown as mean±SD, Welch’s t-test. MFI; mean fluorescence intensity. ( d ) Flow cytometry of relative abundances of Ki67+ tumor cells (mT neg CD45 neg ), total immune cells (mT+ CD45 + ), and stromal cells (mT+ CD45 neg ) in control vs. NOTCH4-treated tumors (n=6 replicates). Data are shown as mean ±SD, 2-way ANOVA test, Bonferroni-corrected. ( e ) Immunofluorescence staining of CD31, IBA1, and Ki67 in tumor transplants from (a). Scale bars, 100μm. ( f ) Primary tumor growth (left) and lung metastasis (right) of metastatic TNBC PDX cells (BR18) in NSG mice following NOTCH4 and IgG treatment. Mice were engrafted with 50K cells and treated as in ( c). Growth kinetics and lung dissemination were evaluated by bioluminescence.

Article Snippet: Antibodies against ARG1 (#79404), Notch Activated Targets Antibody Sampler Kit (#68309), phospho-SMAD2/3 (#8685), CSF-1R (#3152), phospho-AKT Ser473 (#4060), AKT (#9272) and phospho-ERK (#9101) were purchased from Cell Signaling.

Techniques: Western Blot, Inhibition, Isolation, Control, Flow Cytometry, Expressing, Protease Inhibitor, Cell Culture, Labeling, Fluorescence, Immunofluorescence, Staining

Journal: Cell Reports Medicine

Article Title: Light-driven thermotropic hydrogel with nanomodulator enables lipid intervention and cavity delivery for postsurgical tumor therapy

doi: 10.1016/j.xcrm.2026.102880

Figure Lengend Snippet: The effects of LA on the phenotypic characteristics of RAW264.7 cells and regulation of Ra on their phenotypic expression (A–C) The Kaplan-Meier survival curves of ABCA1, ABCG1, and PLIN4 in TCGA-SKCM. (D–J) The transcriptomic analysis of RAW264.7 cells with different treatments: (D) the volcano plots of LA group (culturing for three generations with LA) compared with control group (culturing for three generations without LA), and both the blue and red dots represented SDEGs (|log 2 FC|≥1 and q < 0.05); (E and F) GO and KEGG enrichment analysis of lipid synthesis/metabolism genes in LA group compared with control group; (G) the lipid synthesis/metabolism gene heatmap of control and LA groups; (H and I) GO and KEGG enrichment analysis of immune-related genes in LA group compared with control group; (J) the immune-related gene heatmap of control and LA groups ( n = 3). (K and L) The flow cytometry assay of LD levels in CD206 − RAW264.7 and CD206 + RAW264.7 ( n = 3). (M) The strategy of lipid droplet metabolism to regulate RAW264.7 phenotype. (N and O) The flow cytometry assay of LD levels in RAW264.7 treated with different Ra concentrations ( n = 3). (P and Q) Perilipin 2, iNOS, and Arg-1 expression levels of RAW264.7 treated with Ra ( n = 3). Data are represented as mean ± SD. p values were determined by one-way ANOVA with Tukey’s test for (L), (N), and (Q); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also .

Article Snippet: Anti-mouse Arg-1 Antibody , Abclonal , Cat#A4923; RRID:AB_2863390.

Techniques: Expressing, Control, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Light-driven thermotropic hydrogel with nanomodulator enables lipid intervention and cavity delivery for postsurgical tumor therapy

doi: 10.1016/j.xcrm.2026.102880

Figure Lengend Snippet: The modulation of R@LOX on the polarization of macrophages (A) Representative flow cytometry dot plots. (B) The proportion of M1/M2 in BMDMs ( n = 3). (C–F) The relative mRNA expression of (C) CD86, (D) NOS2, (E) CD206, and (F) Arg-1 in BMDMs ( n = 3). (G–K) The transcriptomic analysis results of RAW264.7 cells: (G) the volcano plots of R@LOX group (culturing with R@LOX and LA) compared with control group (culturing with only LA), and both the blue and red dots represented SDEGs (|log 2 FC|≥1 and q < 0.05); (H and I) GO and KEGG enrichment analysis of SDEGs in R@LOX group compared with control group; (J and K) the heatmap of control and R@LOX group genes. Data are represented as mean ± SD. p values were determined by one-way ANOVA with Tukey’s test for (C), (D), (E), and (F) and unpaired two-tailed Student’s t test for (B). n.s., not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. See also .

Article Snippet: Anti-mouse Arg-1 Antibody , Abclonal , Cat#A4923; RRID:AB_2863390.

Techniques: Flow Cytometry, Expressing, Control, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Light-driven thermotropic hydrogel with nanomodulator enables lipid intervention and cavity delivery for postsurgical tumor therapy

doi: 10.1016/j.xcrm.2026.102880

Figure Lengend Snippet: In vivo immune activation mechanism of LTG@RL against postsurgical melanoma recurrence (A–D) Relative mRNA expressions of (A) TNF-α, (B) IFN-γ, (C) Arg-1, (D) NOS2 in tumor tissues ( n = 3). (E–S) Flow cytometric analysis of (E and F) T cells, (G and H) CD8 + T cells, (I and J) CD86 + TAM, (K and L) CD86 + NOS2 + TAM, (M and N) CD206 + TAM, (O and P) CD206 + Arg-1 + TAM, proportion of (Q) M1/M2 and (R and S) PD-L2 + TAM in tumors ( n = 3, the unstained control, compensation control, and viability dye control were employed). (T) Flow cytometry assay of LD levels in CD11b + F4/80 + cells in tumors ( n = 5). Data are represented as mean ± SD. p values were determined by one-way ANOVA with Tukey’s test for (A), (B), (C), (D), and (T), and unpaired two-tailed Student’s t test for (F), (H), (J), (L), (N), (P), (Q), and (S); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. See also .

Article Snippet: Anti-mouse Arg-1 Antibody , Abclonal , Cat#A4923; RRID:AB_2863390.

Techniques: In Vivo, Activation Assay, Control, Flow Cytometry, Two Tailed Test