Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: The 3D culture of esophageal organoids in EEM hydrogel. a) Brightfield images of esophageal organoids cultured in various concentrations of EEM hydrogels and MAT at day 9. b) Quantification of organoid formation efficiency in EEM hydrogels, relative to MAT ( n = 3, one-way ANOVA with Tukey's multiple comparisons test; ∗∗ p < 0.01; ns, non-significant difference with p > 0.05). c) qRT-PCR analysis to compare gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogels and MAT at day 9 ( n = 4, one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05). d) Relative gene expression of basal layer markers ( Sox2 and Krt14 ) and suprabasal layer markers ( Krt13 and Krt4 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at day 9 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT). e) Morphology of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 9 days. f) Immunofluorescent staining for basal layer markers (CK14, SOX2, and p63), suprabasal layer markers (CK13 and CK4), F-actin, a cell-cell adhesion marker (ECAD), and a proliferation marker (Ki67) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days. g) Quantification of CK14 and CK13 positive area in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days, based on immunofluorescence-stained images ( n = 4; unpaired two-tailed t -test; ∗ p < 0.05 versus MAT; ns, non-significant difference with p > 0.05). h) Live/Dead staining to assess the viability of organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 11 days. i) Representative brightfield images at passage 0, 2, 4, and 6 of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT. Total culture days are given in parentheses. j) Comparison of gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at passage 0 and 6 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT at each passage).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Quantitative RT-PCR, Gene Expression, Marker, Two Tailed Test, Staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: LT-NPs-NIR protects TSPCs against oxidative stress-induced senescence and preserves tenogenic phenotype. (A–D) Immunofluorescence staining for DNA damage (γ-H2AX), proliferation (Ki67), and senescence markers (P16, P53). (E–G) Assessment of stemness (SOX2) and tenogenic differentiation markers (SCX, COL1). (H) Quantitative analysis of the indicated markers. (I) qRT-PCR analysis of SASP-related inflammatory mediators (IL-1β, CXCL10) and matrix-degrading enzymes (MMP3, MMP13). (J) Schematic illustrating the mechanism of ROS scavenging and SASP inhibition. Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Snt: senescent cells; Yng: young cells.

Article Snippet: After washing, cells were incubated with primary antibodies against Ki67 (ab15580, Abcam), Phosphorylated Histone H2AX (γ-H2AX) (ab81299, Abcam), SOX2 (sc-365964, Santa Cruz), Type I Collagen (COL1) (ab138492, Abcam), tenomodulin (TNMD) (ab203676, Abcam; sc-51813, Santa Cruz), Scleraxis (SCX) (sc-518082, Santa Cruz), IRF3 (ab68481, Abcam), Transcription Factor p65/RELA (P65) (A22331, Abclonal), Cyclin-Dependent Kinase Inhibitor 2A (p16INK4a) (P16) (sc-1661, Santa Cruz), P53 (10442-1-AP, Proteintech), Inducible Nitric Oxide Synthase (iNOS) (ab178945, Abcam), Arginase-1(Arg-1) (ab96183, Abcam), HSP70 (sc-32239, Santa Cruz), IL-6 (ab233706, Abcam), Matrix Metalloproteinase 13 (MMP13) (ab39012, Abcam), Double-stranded DNA (dsDNA) Marker (sc-58749, Santa Cruz), and Translocase of Outer Mitochondrial Membrane 20 (TOMM20) (11802-1-AP, Proteintech).

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Inhibition

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: LT-NPs-NIR modulate macrophage polarization and enhance TSPC-mediated tenogenic repair in a Transwell co-culture system. (A) Schematic of the Transwell co-culture setup. (B) SA-β-gal staining of macrophages. (C–F) Immunofluorescence of TSPCs for (C) P16, (D) SOX2, (E) SCX, and (F) Tenomodulin (TNMD) with F-actin. (G) Quantification of P16, SOX2, SCX, and TNMD levels. (H) Proposed mechanism: LT-NPs-NIR promote an M1-to-M2 macrophage shift and regulate TSPC senescence/stemness balance to favor tenogenic repair, potentially via STING/NF-κB signaling. Scale bars: 100 μm (B); 50 μm (C–E); 100 μm (F). Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After washing, cells were incubated with primary antibodies against Ki67 (ab15580, Abcam), Phosphorylated Histone H2AX (γ-H2AX) (ab81299, Abcam), SOX2 (sc-365964, Santa Cruz), Type I Collagen (COL1) (ab138492, Abcam), tenomodulin (TNMD) (ab203676, Abcam; sc-51813, Santa Cruz), Scleraxis (SCX) (sc-518082, Santa Cruz), IRF3 (ab68481, Abcam), Transcription Factor p65/RELA (P65) (A22331, Abclonal), Cyclin-Dependent Kinase Inhibitor 2A (p16INK4a) (P16) (sc-1661, Santa Cruz), P53 (10442-1-AP, Proteintech), Inducible Nitric Oxide Synthase (iNOS) (ab178945, Abcam), Arginase-1(Arg-1) (ab96183, Abcam), HSP70 (sc-32239, Santa Cruz), IL-6 (ab233706, Abcam), Matrix Metalloproteinase 13 (MMP13) (ab39012, Abcam), Double-stranded DNA (dsDNA) Marker (sc-58749, Santa Cruz), and Translocase of Outer Mitochondrial Membrane 20 (TOMM20) (11802-1-AP, Proteintech).

Techniques: Co-Culture Assay, Staining, Immunofluorescence

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: 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, including p-STAT3 (cat. no. 4113, Cell Signaling Technology), MCL1 (cat. no. 16225-1-AP), MMP2 (cat. no. 10373-2-AP) and SOX2 (cat. no. 11064-1-AP; all Proteintech Group, Inc.), were diluted 1:100 in antibody diluent (cat. no. PR30016; Proteintech Group, Inc.) and applied at 4°C overnight.

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