Journal: Fundamental Research

Article Title: Engineered In-Situ-Forming Biomimetic Hydrogel with Self-Regulated Immunostimulatory Capacity Promotes Postoperative Tumor Treatment

doi: 10.1016/j.fmre.2023.02.029

Figure Lengend Snippet: Fig. 3. Ferroptosis-inducing and immunostimulatory capabilities of the biomimetic hydrogel. (a) Schematic diagram for the tumor cell/immune cell co- incubation system in transwell plates. The tumor cells (B16F10, 4T1) or immune cells were inoculated in the bottom chamber of the 24-well transwell culture plate, while the hydrogel soaking solution was placed in the upper chamber. (b) Changes of GPX4 activity in B16F10 cells after different treatments. (I) Control, (II) Gel, (III) RSL3, (IV) Gel@RSL3 (n = 4). (c) Flow cytometric analysis on the lipid ROS levels in B16F10 cells after different treatments. (d) CLSM imaging of lipid ROS generation in B16F10 cells after different treatments. Higher green fluorescence intensity indicates greater lipid ROS production. (e) Quantitative fluorescence analysis of lipid ROS levels in panel D (n = 4). (f) Flow cytometric analysis on the hydrogel-mediated ferroptosis levels of B16F10 cells after different treatments. (g) ATP levels in the supernatants of cell culture after different treatments. (I) Control, (II) Gel, (III) RSL3, (IV) Gel@RSL3 (n = 4). (h) CLSM imaging of CRT expression in B16F10 cells after different treatments. Stronger red fluorescence indicates higher expression levels. (i) Quantitative fluorescence analysis of CRT expression levels in panel H (n = 4). (j) CLSM imaging of cellular HMGB1 abundance after different treatments. Lower red fluorescence indicates greater HMGB1 release into the extracellular compartment. (k) Quantitative fluorescence analysis of HMGB1 release in panel J (n = 4). (l) Flow cytometric analysis on the treatment-induced maturation of BMDCs. (m) Flow cytometric analysis on the activation status of macrophages by monitoring the CD80 expression levels. ∗ indicates significance at P < 0.05, ∗ ∗ indicates significance at P < 0.01.

Article Snippet: RT, HMGB1, and SLC7A11 were supplied by Proteintech (Wuhan, hina).

Techniques: Incubation, Activity Assay, Control, Imaging, Cell Culture, Expressing, Activation Assay

Journal: Fundamental Research

Article Title: Engineered In-Situ-Forming Biomimetic Hydrogel with Self-Regulated Immunostimulatory Capacity Promotes Postoperative Tumor Treatment

doi: 10.1016/j.fmre.2023.02.029

Figure Lengend Snippet: Fig. 4. Gel@RSL3 + GM-CSF + aPD-L1 activates immune response in vitro. (a-d) Flow cytometric analysis of the activation status of DCs (CD11c + /MHC II + ), M1 macrophages (F4/80 + /CD80 + ) and T cells (CD8 + /CD3 + and CD8a + /IFN- 𝛾+ ) in the co-incubation system of splenic immune cells and B16F10 cells after treatment with (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3 and (V) Gel@RSL3 + GM-CSF (n = 4). (e) Secretion levels of immunostimulatory cytokines including IFN- 𝛾, TNF- 𝛼and antitumor effector molecule GzmB in the supernatant from the co-culture system after different treatments (n = 4). (f) PD-L1 expression in tumor cells with after the hydrogel-mediated ferroptosis-immunotherapy. Group set-up for panel e-f: (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3 and (V) Gel@RSL3 + GM- CSF). (g-h) Flow cytometric analysis of the expression levels of effector T cell marker CD4 + /CD8 + and CD8a + /IFN- 𝛾+ in T cells co-incubated with B16F10 cells after treatment with (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3, (V) Gel@RSL3 + GM-CSF and (VI) Gel@RSL3 + GM-CSF + aPD-L1. (i) Secretion levels of immunostimulatory cytokines including IFN- 𝛾, TNF- 𝛼and antitumor effector molecule GzmB in the supernatant from the co-culture system after different treatments (n = 4). (j) Evaluation on the GSH levels in B16F10 cells after different treatments (n = 4). (k) Western blot analysis of the expression level of CRT, HMGB1 and SLC7A11 in different groups. (l) Flow cytometric analysis on the lipid ROS levels in B16F10 cells after different treatments. (m) MDA levels in B16F10 cells after different treatments (n = 4). Group set-up for panel I-M: (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3, (V) Gel@RSL3 + GM-CSF and (VI) Gel@RSL3 + GM-CSF + aPD- L1). (n) Flow cytometric analysis on the death rate of B16F10 cells after different treatments, including (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3, (V) Gel@RSL3 + GM-CSF and (VI) Gel@RSL3 + GM-CSF + aPD-L1. ∗ indicates significance at P < 0.05, ∗ ∗ indicates significance at P < 0.01, ∗ ∗ ∗ indicates significance at P < 0.001, ∗ ∗ ∗ ∗ indicates significance at P < 0.0001.

Article Snippet: RT, HMGB1, and SLC7A11 were supplied by Proteintech (Wuhan, hina).

Techniques: In Vitro, Activation Assay, Incubation, Control, Co-Culture Assay, Expressing, Marker, Western Blot

Journal: Fundamental Research

Article Title: Engineered In-Situ-Forming Biomimetic Hydrogel with Self-Regulated Immunostimulatory Capacity Promotes Postoperative Tumor Treatment

doi: 10.1016/j.fmre.2023.02.029

Figure Lengend Snippet: Fig. 5. Antitumor effect of biomimetic hydrogel in vivo. (a) Schematic illustration of the treatment scheme of the B16F10-luc tumor-bearing mice (n = 7). (b) Treatment procedures on the tumor-bearing mice. (c) In vivo bioluminescence images of B16F10-luc tumor-bearing mice throughout the treatment period with (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3, (V) Gel@RSL3 + GM-CSF and (VI) Gel@RSL3 + GM-CSF-aPD-L1. (d) Tumor size changes during the incubation period after different treatments. (e) Survival analysis of mice after different treatments. (f) Body weight changes after treatment with different samples. (g) Evaluation on the GPX4 activity in tumor tissues after different treatments (n = 4). (h) Western blot analysis on the expression of CRT, HMGB1 and SLC7A11 in B16F10-luc tumors after treatment with (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3, (V) Gel@RSL3 + GM-CSF and (VI) Gel@RSL3 + GM-CSF-aPD-L1. (i) MDA levels in tumor tissue after different treatments (n = 4). (j) H&E and TUNEL staining of tumor tissue samples after treatment (I) Control, (II) Gel, (III) Gel@GM-CSF, (IV) Gel@RSL3, (V) Gel@RSL3 + GM-CSF and (VI) Gel@RSL3 + GM-CSF-aPD-L1. ∗ indicates significance at P < 0.05, ∗ ∗ indicates significance at P < 0.01, ∗ ∗ ∗ indicates significance at P < 0.001.

Article Snippet: RT, HMGB1, and SLC7A11 were supplied by Proteintech (Wuhan, hina).

Techniques: In Vivo, Control, Incubation, Activity Assay, Western Blot, Expressing, TUNEL Assay, Staining

Journal: Food Science and Human Wellness

Article Title: Sucrose-free hawthorn leathers formulated with fructooligosaccharides and xylooligosaccharides ameliorate high-fat diet induced inflammation, glucose and lipid metabolism in liver of mice

doi: 10.1016/j.fshw.2022.03.033

Figure Lengend Snippet: Fig. 4 Effect of hawthorn leather treatment on expression levels of inflammatory cytokines and NF-κB signaling pathway in the liver tissue of mice. mRNA expression level of (A) IL-1β, (B) Nos2, (C) Cox2, (D) TLR4, (E) MyD88, and (F) NF-κB; (G) Images of western blotting; Protein expression ratio of (H) MyD88 to GAPDH and (I) NF-κB to GAPDH. Data are expressed as mean ± SD (8 mice/group). Values of each group with different letters over the bars are significantly different (P < 0.05) by analysis of one-way ANOVA followed by Tukey’s multiple range test.

Article Snippet: Then, the membrane was incubated with specific primary antibody (1 : 1 000) against MyD88 (CST, cat. number 4283S), NF-κB (p65) (CST, cat. number 8242S), PI3K (p110β) (Proteintech, cat. number 20584-1-AP), AKT (Proteintech, cat. number 10176-2-AP), Thr308 phosphorylated AKT (p-AKT) (CST, cat. number 13038T), mTOR (Proteintech, cat. number 20657-1-AP), Ser2448 phosphorylated mTOR (p-mTOR) (CST, cat. number 5536T), PPAR-γ (CST, cat. number 2435T), FASN (CST, cat. number 3180T), and HSL (CST, cat. number 18381T) overnight at 4 °C, respectively.

Techniques: Expressing, Western Blot

Journal: Journal of Functional Foods

Article Title: Oxyresveratrol from mulberry (Morus alba L.) ameliorates post-inflammatory hyperpigmentation in vitro by anti-melanogenesis, inhibiting melanosome transfer, and providing photoprotection

doi: 10.1016/j.jff.2024.106557

Figure Lengend Snippet: Fig. 4. The inhibitory effects of OXY, RSV, and GLA on melanin transport in human melanocytes. (a) 15.67 μg/mL OXY, RSV, and GLA were cultured in human melanocytes for 48 h. The mRNA levels of melanin transport genes RAC1, RAB27A, and CDC42 were determined by qRT-PCR. (b) The protein expression level of melanin transport genes RAC1, RAB27A, and CDC42 was determined by western blotting using GAPDH as a loading control. Values are means ± SD of three in dependent experiments run in triplicate. *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared to the control.

Article Snippet: Primary antibodies against CDC42 (10155–1- AP), RAB27A (17817–1-AP), RAC1 (24072–1-AP), and GAPDH (60004–1-Ig), beta-actin polyclonal antibody (β-actin, 20536–1-AP) were purchased from ProteinTech (Wuhan, China).

Techniques: Cell Culture, Quantitative RT-PCR, Expressing, Western Blot, Control

Journal: Journal of Functional Foods

Article Title: Oxyresveratrol from mulberry (Morus alba L.) ameliorates post-inflammatory hyperpigmentation in vitro by anti-melanogenesis, inhibiting melanosome transfer, and providing photoprotection

doi: 10.1016/j.jff.2024.106557

Figure Lengend Snippet: Fig. 5. Interaction of OXY with RAB27A, RAC1, and CDC42. The molecular docking results between OXY and RAB27A (a), RAC1 (b), and CDC42 (c) were visualized by PyMOL. The protein models of RAB27A (ID: 3BC1), RAC1 (ID: 3TH5), and CDC42 (ID: 1E0A) were downloaded from the RCSB Protein Data Bank. The ligand, metal ion and protein were labeled in red, yellow, and blue, respectively. The hydrogen bonds between the ligand and protein were repre sented by dotted yellow lines. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Primary antibodies against CDC42 (10155–1- AP), RAB27A (17817–1-AP), RAC1 (24072–1-AP), and GAPDH (60004–1-Ig), beta-actin polyclonal antibody (β-actin, 20536–1-AP) were purchased from ProteinTech (Wuhan, China).

Techniques: Labeling