Journal: International Journal of Molecular Medicine

Article Title: Deciphering the CAF-LCN2 axis: Key to overcoming anti-PD-L1 immunotherapy resistance in lung cancer

doi: 10.3892/ijmm.2026.5735

Figure Lengend Snippet: Influence of LCN2 on the growth of lung cancer in mice. (A) Simplified flowchart outlining the establishment of an anti-PD-L1 immunotherapy tolerance model in mice with lung cancer. (B) Western blot analysis of LCN2, COL1A1 and THY1 expression in tumor tissues from drug-resistant and drug-sensitive mice. (C) Immunohistochemical detection of LCN2, COL1A1 and THY1 expression in tumor tissues from drug-resistant and drug-sensitive mice (scale bar, 100 µm). (D) Establishment of a model for anti-PD-L1 immunotherapy in LCN2-silenced mice. (E) Evaluation of LCN2 mRNA expression in tumor tissues after LCN2 gene intervention in mice. (F) Assessment of LCN2 protein expression in tumor tissues following LCN2 gene intervention in mice. (G) Tumor size in LCN2 gene intervention mice (n=10 in each group). *P<0.05, **P<0.01 and ***P<0.001. Unpaired two-tailed Student's t-test was used for (B) and (C) and one-way ANOVA with Tukey's multiple comparisons test was used for (E) and (G). LCN2, lipocalin 2; PD-L1, programmed death-ligand 1; sh-, short hairpin; NC, negative control; COL1A1, collagen α-1(I) chain; THY1, Thy-1 membrane glycoprotein; i.p., intraperitoneal.

Article Snippet: Following permeabilization, cells were incubated overnight at 4°C with the following primary antibodies: Rabbit anti-collagen α-1(I) chain (COL1A1) (cat. no. PA5-101300; 1:250; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-Cytokeratin (cat. no. ab53280; 1:500; Abcam) and mouse anti-CD31 (cat. no. sc-376764; 1:50; Santa Cruz Biotechnology, Inc.).

Techniques: Western Blot, Expressing, Immunohistochemical staining, Two Tailed Test, Negative Control, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Linked to Reepithelialization of Human Skin Wounds

doi: 10.3390/ijms232415746

Figure Lengend Snippet: Box plot of injury-induced DEGs in wound tissue (blue symbols) vs. adjacent normal skin (green symbols) tissue of 4-day-old deroofed suction blisters ( n = 17) arranged in descending FC order. Boxes represent the 25th–75th percentiles, whiskers represent the 5th–95th percentiles, and the horizontal dashed lines within the boxes indicate the median values. The y-axis shows log2-transformed expression values. FC, fold change.

Article Snippet: The tissue sections were subjected to heat-induced epitope retrieval pretreatment using EnVisionTM FLEX Target Retrieval Solution High pH (GV804, Dako Omnis, Agilent) for TIMP-1 or EnVisionTM FLEX Target Retrieval Solution Low pH (GV805, Dako Omnis, Agilent) for collagen I for 30 min, followed by incubation with rabbit monoclonal antibodies against TIMP-1 (clone EPR18352, 1:500, ab211926, Abcam, Cambridge, UK) or collagen α1 (I) (E8F4L, 1:100, #72026, Cell Signaling, Danvers, MA, USA) for 30 min at 32 °C.

Techniques: Transformation Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Linked to Reepithelialization of Human Skin Wounds

doi: 10.3390/ijms232415746

Figure Lengend Snippet: Effects of IL-6 (30 ng/mL) and IL-1β (1 ng/mL) treatment of NHEKs ( A , B ) and NHDFs ( C , D ) on MMP1 and TIMP1 mRNA ( A , C ) and protein media levels of MMP-1 and TIMP-1 ( B , D ). mRNA levels were determined by RT–qPCR and normalized to RPLP0 , expressed as FC relative to control-treated NHEKs and NHDFs. The geometric mean ± back-transformed SEM of 6 replicates is shown ( A , C ). MMP-1 and TIMP-1 protein levels were determined by ELISA. The mean ± SEM of 6 replicates is shown ( B , D ). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control.

Article Snippet: The tissue sections were subjected to heat-induced epitope retrieval pretreatment using EnVisionTM FLEX Target Retrieval Solution High pH (GV804, Dako Omnis, Agilent) for TIMP-1 or EnVisionTM FLEX Target Retrieval Solution Low pH (GV805, Dako Omnis, Agilent) for collagen I for 30 min, followed by incubation with rabbit monoclonal antibodies against TIMP-1 (clone EPR18352, 1:500, ab211926, Abcam, Cambridge, UK) or collagen α1 (I) (E8F4L, 1:100, #72026, Cell Signaling, Danvers, MA, USA) for 30 min at 32 °C.

Techniques: Quantitative RT-PCR, Control, Transformation Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Linked to Reepithelialization of Human Skin Wounds

doi: 10.3390/ijms232415746

Figure Lengend Snippet: TIMP-1 ( A , B ) and collagen α1 (I) ( C , D ) immunohistochemical analysis of a 4-day-old wound ( A , B , D ) with adjacent skin ( C ). ( A ) Fibroblasts (solid arrowheads) below neoepidermis and ( B ) endothelial cell (outline arrowhead) of small blood vessel show positive immunoreactivity of TIMP-1 in upper dermis. ( D ) Arrow indicates the tip of the migrating neoepidermal tongue.

Article Snippet: The tissue sections were subjected to heat-induced epitope retrieval pretreatment using EnVisionTM FLEX Target Retrieval Solution High pH (GV804, Dako Omnis, Agilent) for TIMP-1 or EnVisionTM FLEX Target Retrieval Solution Low pH (GV805, Dako Omnis, Agilent) for collagen I for 30 min, followed by incubation with rabbit monoclonal antibodies against TIMP-1 (clone EPR18352, 1:500, ab211926, Abcam, Cambridge, UK) or collagen α1 (I) (E8F4L, 1:100, #72026, Cell Signaling, Danvers, MA, USA) for 30 min at 32 °C.

Techniques: Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Linked to Reepithelialization of Human Skin Wounds

doi: 10.3390/ijms232415746

Figure Lengend Snippet: Macrodissection of FFPE tissue section of a day-4 suction blister wound into central wound (1 piece) including neoepidermis and adjoining nonwounded normal skin including epidermis (2 pieces). Reepithelialization was calculated using the following formula: Neoepidermis (mm)/Wound length (mm) × 100%. Total RNA was extracted, and the extracts were subjected to nCounter mRNA analysis. Epidermal compartments are black and dermal compartments blue. Scale bar, 1 mm.

Article Snippet: The tissue sections were subjected to heat-induced epitope retrieval pretreatment using EnVisionTM FLEX Target Retrieval Solution High pH (GV804, Dako Omnis, Agilent) for TIMP-1 or EnVisionTM FLEX Target Retrieval Solution Low pH (GV805, Dako Omnis, Agilent) for collagen I for 30 min, followed by incubation with rabbit monoclonal antibodies against TIMP-1 (clone EPR18352, 1:500, ab211926, Abcam, Cambridge, UK) or collagen α1 (I) (E8F4L, 1:100, #72026, Cell Signaling, Danvers, MA, USA) for 30 min at 32 °C.

Techniques: