Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits STAT3 signaling activation in hepatocellular carcinoma cells. The protein levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The protein levels of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous tumor tissue were detected by immunohistochemical staining. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; p, phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2, SRY-box transcription factor 2.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Activation Assay, Western Blot, Immunohistochemical staining, Staining, Negative Control, Sequencing

Journal: iScience

Article Title: Changes in the microglial phenotype drive neuroinflammation independent of systemic inflammation in the acute stage of heatstroke

doi: 10.1016/j.isci.2026.115254

Figure Lengend Snippet: Time course of functional damage to the intestinal barrier and blood-brain barrier during the acute stage of heatstroke The mice were exposed to 41.2 ± 0.5°C ambient temperature until their rectal temperature reached 42.4°C, and then they were allowed to recover at an ambient temperature of 25 ± 0.5°C for the indicated times. (A) The concentration of FD-4 in serum was measured after its oral administration. (B) Serum endotoxin levels. (C) EB was injected into the mice via the tail vein after the onset of heatstroke, and representative images of EB leakage was shown. (D) Quantitative analysis of EB concentrations in brain tissues. (E) Expression of MMP-2 in the cerebral cortex at 1 h, 6 h, and 24 h post heatstroke. (F) Expression of MMP-9 in the cerebral cortex at 1 h, 6 h, and 24 h post heatstroke. The data are expressed as the mean ± SEM of three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, vs. Con.

Article Snippet: The membrane was further incubated overnight at 4°C with primary antibodies against MMP9 (TA326652; ORIGENE, USA), MMP2 (TA326260; ORIGENE, USA) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 2118L; CST, USA).

Techniques: Functional Assay, Concentration Assay, Injection, Expressing

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates the intestinal PPAR-γ and downregulates the MMP-9/2 expression. (A) The degree of the node between the HDCA and the intersection target. (B) Molecular docking analysis between HDCA and PPAR-γ. (C) Molecular docking analysis between HDCA and MMP-9. (D) Molecular docking analysis between HDCA and MMP-2. (E-G) Relative expression of mRNA of ppar-γ, mmp9 and mmp2 in the distal ileum. (n = 6) . (H) Representative protein immunoblots in distal ileum. (I-K) Relative expression of PPAR-γ, MMP-9, MMP-2 (n = 3–4). (L) Representative immunohistochemical staining and quantitative analysis of MMP-2+ (M), PPAR-γ+ (N) and MMP-9+ (O) cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (E–G, I–K, M–O). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates PPAR-γ expression and reduces MMP-9/2 expression in the spinal cord. (A–C) The relative expression of mRNA in spinal cord of ppar-γ, mmp9 and mmp2 . (n = 6) . (D–G) Representative immunoblots of proteins and relative expression of PPAR-γ, MMP-2, MMP-9 (n = 4). (H–J) Immunofluorescence staining of PPAR-γ, MMP-9, MMP-2 in spinal cord (scale, 100 μm). (n = 3). (K) Heatmap of Correlation Analysis. * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (A–D, F, G). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: Fxr knock down abolished the protective effect of HDCA in neuropathic pain. (A) Immunofluorescence staining showing FXR co-localization in spinal cord. (scale, 25 μm). (B) Immunofluorescence staining demonstrating MMP-2 colocalization in spinal cord. (scale, 25 μm). (C) Immunofluorescence staining showing PPAR-γ colocalization in spinal cord. (scale, 25 μm). (D) Immunofluorescence staining showing MMP-9 colocalization in spinal cord. (scale, 25 μm). (E) Immunofluorescence staining of FXR and MMP-2 colocalization in spinal cord. (scale, 25 μm). (F) Representative immunoblots of protein expression in the spinal cord from Fxr -/- mice. (G) Relative expression of PPAR-γ, MMP-2 and MMP-9 (n = 4). (H) Representative immunoblots for the proteins in spinal cord following INT-747 treatment. (I) PWT of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (J) PWL of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (K) Relative expression of PPAR-γ, MMP-2 and MMP-9 following INT-747 treatment. (n = 4). (L, M) Representative immunoblots for the proteins in spinal cord and relative expression of PPAR-γ, MMP-2 and MMP-9 in Fxr -/- mice following HDCA treatment. (n = 4). * P < 0.05, ** P < 0.01, ns by unpaired Student's t‐test (G, M). ns by Two-way repeated ANOVA with post hoc Bonferroni’s test (I and J), * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (K). Data are represented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Knockdown, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates the intestinal PPAR-γ and downregulates the MMP-9/2 expression. (A) The degree of the node between the HDCA and the intersection target. (B) Molecular docking analysis between HDCA and PPAR-γ. (C) Molecular docking analysis between HDCA and MMP-9. (D) Molecular docking analysis between HDCA and MMP-2. (E-G) Relative expression of mRNA of ppar-γ, mmp9 and mmp2 in the distal ileum. (n = 6) . (H) Representative protein immunoblots in distal ileum. (I-K) Relative expression of PPAR-γ, MMP-9, MMP-2 (n = 3–4). (L) Representative immunohistochemical staining and quantitative analysis of MMP-2+ (M), PPAR-γ+ (N) and MMP-9+ (O) cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (E–G, I–K, M–O). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates PPAR-γ expression and reduces MMP-9/2 expression in the spinal cord. (A–C) The relative expression of mRNA in spinal cord of ppar-γ, mmp9 and mmp2 . (n = 6) . (D–G) Representative immunoblots of proteins and relative expression of PPAR-γ, MMP-2, MMP-9 (n = 4). (H–J) Immunofluorescence staining of PPAR-γ, MMP-9, MMP-2 in spinal cord (scale, 100 μm). (n = 3). (K) Heatmap of Correlation Analysis. * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (A–D, F, G). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: Fxr knock down abolished the protective effect of HDCA in neuropathic pain. (A) Immunofluorescence staining showing FXR co-localization in spinal cord. (scale, 25 μm). (B) Immunofluorescence staining demonstrating MMP-2 colocalization in spinal cord. (scale, 25 μm). (C) Immunofluorescence staining showing PPAR-γ colocalization in spinal cord. (scale, 25 μm). (D) Immunofluorescence staining showing MMP-9 colocalization in spinal cord. (scale, 25 μm). (E) Immunofluorescence staining of FXR and MMP-2 colocalization in spinal cord. (scale, 25 μm). (F) Representative immunoblots of protein expression in the spinal cord from Fxr -/- mice. (G) Relative expression of PPAR-γ, MMP-2 and MMP-9 (n = 4). (H) Representative immunoblots for the proteins in spinal cord following INT-747 treatment. (I) PWT of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (J) PWL of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (K) Relative expression of PPAR-γ, MMP-2 and MMP-9 following INT-747 treatment. (n = 4). (L, M) Representative immunoblots for the proteins in spinal cord and relative expression of PPAR-γ, MMP-2 and MMP-9 in Fxr -/- mice following HDCA treatment. (n = 4). * P < 0.05, ** P < 0.01, ns by unpaired Student's t‐test (G, M). ns by Two-way repeated ANOVA with post hoc Bonferroni’s test (I and J), * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (K). Data are represented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Knockdown, Immunofluorescence, Staining, Western Blot, Expressing