Journal: Bioactive Materials

Article Title: TLR2-PI3K/Akt mediated microbe-mimetic priming boosts the therapeutic paracrine function of GelMA-Encapsulated MSCs for diabetic wound regeneration

doi: 10.1016/j.bioactmat.2025.10.007

Figure Lengend Snippet: TLR2-mediated PI3K/Akt signaling drives MSCs functional enhancement by PGN and LTA. (A) Knockdown efficiency of si-TLR2. (B, C) Expression of pathway-related proteins after TLR2 knockout MSCs (siTLR2) and PGN/LTA rescue (B), with quantification (C) (n = 3). (D) mTOR gene expression after TLR2 knockout and rescue. (E) RT-qPCR (n = 3) validation of functional changes of MSCs in angiogenesis and inhibit inflammation after TLR2 knockout. (F, G) Expression of functional proteins after TLR2 knockout MSCs (siTLR2) and PGN/LTA rescue. (H) TLR2 expression after LY294 inhibition pathway. (I) Expression of pathway-related proteins after LY294 and rescue (n = 3). (J) mTOR gene expression after LY294 knockout and rescue. (K) RT-qPCR (n = 3) validation of functional changes after LY294. (L, M) Expression of functional proteins after LY294 and PGN/LTA rescue.

Article Snippet: Bovine serum albumin solutions (5 %) were used to block non-specific antigens for 1.5 h. The membranes were probed with anti-PI3K rabbit monoclonal antibody (1:500, C67E7, Cell siginting), anti-Phospho-Akt (Ser473) rabbit monoclonal antibody (1:500, 9271T, Cell siginting), anti- P13K rabbit monoclonal antibody (1:500, C73F8, Cell siginting), anti-p-PI3K rabbit monoclonal antibody (1:500, ab278545, Abcam) or anti-GAPDH mouse monoclonal antibody (1:1000, 60 004–1-Ig, Proteintech).

Techniques: Functional Assay, Knockdown, Expressing, Knock-Out, Gene Expression, Quantitative RT-PCR, Biomarker Discovery, Inhibition

Journal: International Journal of Molecular Medicine

Article Title: FBXO22 promotes hepatocellular carcinoma progression via paracrine myo-inositol-induced M2-type polarization of macrophages

doi: 10.3892/ijmm.2025.5707

Figure Lengend Snippet: FBXO22-induced myo-inositol release promotes M2 polarization of THP-1 cells. 97H cells were treated with myo-inositol. (A) A Cell Counting Kit-8 assay was performed to assess cell viability. (B and C) Flow cytometry was performed to detect CD86- and CD206-positive THP-1 cells. (D and E) Western blotting was performed to examine p-PI3K and p-AKT levels in THP-1 cells. * P<0.05 and *** P<0.001. p-, phosphorylated.

Article Snippet: The membrane was maintained with 5% non-fat milk for 1 h at room temperature, and incubated with the primary antibodies against the following proteins: PI3K (1:10,000; cat. no. 67071-1-Ig; Proteintech Group, Inc.), phosphorylated (p-) PI3K (1:1,000; cat. no. AF3242; Affinity Biosciences), AKT1 (1:10,000; cat. no. 80457-1-RR; Proteintech Group, Inc.), p-AKT1 (1:5,000; cat. no. 80462-1-RR, Proteintech Group, Inc.), FBXO22 (1:2,000; cat. no. 13606-1-AP; Proteintech Group, Inc.), IMPA1 (1:300; cat. no. 16593-1-AP; Proteintech Group, Inc.), SLC5A3 (1:1,000; cat. no. DF4521; Affinity Biosciences), PTEN (1:5,000; cat. no. 60300-1-Ig; Proteintech Group, Inc.), and NRF2 (1:2,000; cat. no. 16396-1-AP; Proteintech Group, Inc.). β-actin protein level was the internal control.

Techniques: Cell Counting, Flow Cytometry, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: FBXO22 promotes hepatocellular carcinoma progression via paracrine myo-inositol-induced M2-type polarization of macrophages

doi: 10.3892/ijmm.2025.5707

Figure Lengend Snippet: Myo-inositol induces M2 polarization of THP-1 cells via SLC5A3. THP-1 cells were transfected with si- SLC5A3 . (A) Reverse transcription-quantitative PCR and (B) western blotting were performed to detect SLC5A3 expression. THP-1 cells were transfected with si- SLC5A3 , followed by treatment with 10 μ M myo-inositol. (C and D) Flow cytometry was performed to detect CD86-positive and CD206-positive THP-1 cells. (E and F) Western blotting was performed to examine p-PI3K and p-AKT levels in THP-1 cells. * P<0.05, ** P<0.01 and *** P<0.001. p-, phosphorylated.

Article Snippet: The membrane was maintained with 5% non-fat milk for 1 h at room temperature, and incubated with the primary antibodies against the following proteins: PI3K (1:10,000; cat. no. 67071-1-Ig; Proteintech Group, Inc.), phosphorylated (p-) PI3K (1:1,000; cat. no. AF3242; Affinity Biosciences), AKT1 (1:10,000; cat. no. 80457-1-RR; Proteintech Group, Inc.), p-AKT1 (1:5,000; cat. no. 80462-1-RR, Proteintech Group, Inc.), FBXO22 (1:2,000; cat. no. 13606-1-AP; Proteintech Group, Inc.), IMPA1 (1:300; cat. no. 16593-1-AP; Proteintech Group, Inc.), SLC5A3 (1:1,000; cat. no. DF4521; Affinity Biosciences), PTEN (1:5,000; cat. no. 60300-1-Ig; Proteintech Group, Inc.), and NRF2 (1:2,000; cat. no. 16396-1-AP; Proteintech Group, Inc.). β-actin protein level was the internal control.

Techniques: Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Flow Cytometry

Journal: International Journal of Molecular Medicine

Article Title: FBXO22 promotes hepatocellular carcinoma progression via paracrine myo-inositol-induced M2-type polarization of macrophages

doi: 10.3892/ijmm.2025.5707

Figure Lengend Snippet: Myo-inositol promotes HCC tumor growth and induces M2 polarization in vivo. BALB/c nude mice received a subcutaneous injection of 97H cells to induce HCC. The mice were randomly divided into two groups (n=4 each): PBS group and myo-inositol group. (A and B) Tumor growth was monitored every 3 days for ~3 weeks. (C and D) Western blotting was performed to detect p-PI3K and p-AKT levels in tumor tissues. (E and F) IHC staining was performed to detect CD86-positive and CD206-positive cells in tumor tissues. * P<0.05 and ** P<0.01. HCC, hepatocellular carcinoma; p-, phosphorylated.

Article Snippet: The membrane was maintained with 5% non-fat milk for 1 h at room temperature, and incubated with the primary antibodies against the following proteins: PI3K (1:10,000; cat. no. 67071-1-Ig; Proteintech Group, Inc.), phosphorylated (p-) PI3K (1:1,000; cat. no. AF3242; Affinity Biosciences), AKT1 (1:10,000; cat. no. 80457-1-RR; Proteintech Group, Inc.), p-AKT1 (1:5,000; cat. no. 80462-1-RR, Proteintech Group, Inc.), FBXO22 (1:2,000; cat. no. 13606-1-AP; Proteintech Group, Inc.), IMPA1 (1:300; cat. no. 16593-1-AP; Proteintech Group, Inc.), SLC5A3 (1:1,000; cat. no. DF4521; Affinity Biosciences), PTEN (1:5,000; cat. no. 60300-1-Ig; Proteintech Group, Inc.), and NRF2 (1:2,000; cat. no. 16396-1-AP; Proteintech Group, Inc.). β-actin protein level was the internal control.

Techniques: In Vivo, Injection, Western Blot, Immunohistochemistry