Journal: Frontiers in Physiology

Article Title: Molecular mechanism of Danshen injection in treating endometrial fibrosis induced by intrauterine adhesions via the LAMC2-CD44-TGF-β1-SMAD2/3 signaling pathway

doi: 10.3389/fphys.2026.1794215

Figure Lengend Snippet: LPS induced activation of rat uterine fibroblasts. (A) WB detection of LAMC2 expression levels in epithelial cells, (B) Relative expression of LAMC2 in epithelial cells, (C) Level of LAMC2 in cells detected by ELISA in epithelial cells, (D) WB detection of E-cad, N-cad, α-SMA,TGF-β1, SMAD2/3 in fibroblasts, (E) Relative expression of E-cad, N-cad, α-SMA,TGF-β1, SMAD2, SMAD3 in fibroblasts, (F) Immunofluorescence staining of COL1A1 in fibroblasts, (G) Immunofluorescence staining of COL3A1 in fibroblasts, (H) Immunofluorescence staining of α-SMA in fibroblasts, (I) Mean fluorescence intensity of COL1A1, COL3A1 and α-SMA. Compared to model in fibroblasts, * P < 0.05, ** P < 0.01.

Article Snippet: Primary antibodies: CK-19 (1:1000; YM8269, Immunoway, USA), Vimentin (1:1000; YM8324, Immunoway, USA), GAPDH (1:1000; A19056, ABclonal, China), LAMC2 (1:1000; A1869, ABclonal, China), E-cad (1:1000; A20798, ABclonal, China), N-cad (1:500; A0433, ABclonal, China), α-SMA (1:1000; A2319, ABclonal, China), TGF-β1 (1:2000; A15103, ABclonal, China), SMAD2 (1:5000; bsm-52223R, Bioss, China), SMAD3 (1:1000; A16913, ABclonal, China), p- SMAD2 (1:5000; bs-3419R, Bioss, China), p- SMAD3 (1:2000; Ap0727, ABclonal, China), and a secondary antibody (1:2000; AS014, ABclonal, China).

Techniques: Activation Assay, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Fluorescence

Journal: Frontiers in Physiology

Article Title: Molecular mechanism of Danshen injection in treating endometrial fibrosis induced by intrauterine adhesions via the LAMC2-CD44-TGF-β1-SMAD2/3 signaling pathway

doi: 10.3389/fphys.2026.1794215

Figure Lengend Snippet: Danshen injection improves endometrial fibrosis in IUA rats by inhibiting the LAMC2-CD44-TGF-β1- SMAD2/3 signaling pathway. (A) Uterine Weight, (B) Uterine Length, (C) Grading of intrauterine adhesions in each group, (D) Number of uterine glands, (E) Collagen volume fraction, (F) H&E Staining, (G) MASSON Staining, (H) Immunofluorescence staining of COL1A1, (I) Immunofluorescence staining of COL3A1, (J) Immunofluorescence staining of α-SMA, (K) Mean fluorescence intensity of COL1A1, COL3A1 and α-SMA, L: WB detection of LAMC2, TGF-β1, SMAD2, SMAD3, (L) Relative expression of protein LAMC2, TGF-β1, SMAD2, SMAD3. Compared to model, * P < 0.05, ** P < 0.01. For panels A-C, n = 6 independent animals per group; for panels (D–M) , n=3 independent animals per group.

Article Snippet: Primary antibodies: CK-19 (1:1000; YM8269, Immunoway, USA), Vimentin (1:1000; YM8324, Immunoway, USA), GAPDH (1:1000; A19056, ABclonal, China), LAMC2 (1:1000; A1869, ABclonal, China), E-cad (1:1000; A20798, ABclonal, China), N-cad (1:500; A0433, ABclonal, China), α-SMA (1:1000; A2319, ABclonal, China), TGF-β1 (1:2000; A15103, ABclonal, China), SMAD2 (1:5000; bsm-52223R, Bioss, China), SMAD3 (1:1000; A16913, ABclonal, China), p- SMAD2 (1:5000; bs-3419R, Bioss, China), p- SMAD3 (1:2000; Ap0727, ABclonal, China), and a secondary antibody (1:2000; AS014, ABclonal, China).

Techniques: Injection, Staining, Immunofluorescence, Fluorescence, Expressing

Journal: International journal of molecular sciences

Article Title: miR-212-5p Regulates PM 2.5 -Induced Apoptosis by Targeting LAMC2 and LAMA3.

doi: 10.3390/ijms26041761

Figure Lengend Snippet: Figure 3. miR-212-5p negatively regulates LAMC2 and LAMA3: (A) miRanda (v3.3a) and TargetScan (v5.0) projected miR-212-5p target genes displayed; (B) similarity of rno-miR-212-5p and has-miR- 212-5p genes shown; (C) schematic representation of LAMC2-3′ UTR and LAMA3-3′ UTR with miR- 212-5p target prediction and reporter plasmid construction, where red characters indicate binding sites and ”*” characters indicate mutation sites; (D) luciferase activity assessed in 293T cells after co- transfection of LAMC2-WT, LAMC2-MUT, LAMA3-WT, and LAMA3-MUT with miR-212-5p mimics or miR-mimics NC for 48 h; (E) LAMC2-mVenus-WT, LAMC2-mVenus-MUT, LAMA3-mVenus-WT, and LAMA3-mVenus-MUT co-transfected with miR-mimics NC or miR-212-5p in 293T cells (scale bar: 300 µm); (F) RT-PCR assay for miR-212-5p mimics and miR-212-5p inhibitors on LAMC2 and LAMA3 expression after transfection; (G) Western blotting used to identify alterations in the LAMC2 and LAMA3 proteins. The mean ± standard deviation (n = 3) is used to express the data. ns, no significance; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 indicate significance.

Article Snippet: Next, 5% skimmed milk (Beyotime Biotechnology, Shanghai, China) was blocked for 2 h, and the membranes were incubated with specific primary antibodies overnight at 4 ◦C: BAD, BCL-2, caspase-3, LAMC2, p85, AKT, p-AKT, IκBα, β-action (1:1000 dilution, Proteintech, Rosemont, IL, USA), p-IκBα, p65, p-p65 (1:500 dilution, Proteintech, IL, USA), LAMA3, p-p85 (1:500 dilution, Abways, Shanghai, China), and GAPDH (1:7000 dilution, Abways, Shanghai, China).

Techniques: Plasmid Preparation, Binding Assay, Mutagenesis, Luciferase, Activity Assay, Cotransfection, Transfection, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Standard Deviation

Journal: International journal of molecular sciences

Article Title: miR-212-5p Regulates PM 2.5 -Induced Apoptosis by Targeting LAMC2 and LAMA3.

doi: 10.3390/ijms26041761

Figure Lengend Snippet: Figure 4. PM2.5-induced changes in LAMC2 and LAMA3 expression: (A) LAMC2 and LAMA3 gene expression detected with RT-PCR after PM2.5 stimulation; (B) LAMC2 and LAMA3 protein changes detected with Western blotting; (C) silencing efficiency of LAMC2 and LAMA3 after transfection with siRNA−LAMC2 or siRNA−LAMA3 detected with RT-PCR; (D) LAMC2 and LAMA3 protein changes detected with Western blotting. The mean ± standard deviation (n = 3) is used to express the data. ns, no significance; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 indicate significance.

Article Snippet: Next, 5% skimmed milk (Beyotime Biotechnology, Shanghai, China) was blocked for 2 h, and the membranes were incubated with specific primary antibodies overnight at 4 ◦C: BAD, BCL-2, caspase-3, LAMC2, p85, AKT, p-AKT, IκBα, β-action (1:1000 dilution, Proteintech, Rosemont, IL, USA), p-IκBα, p65, p-p65 (1:500 dilution, Proteintech, IL, USA), LAMA3, p-p85 (1:500 dilution, Abways, Shanghai, China), and GAPDH (1:7000 dilution, Abways, Shanghai, China).

Techniques: Expressing, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Standard Deviation

Journal: International journal of molecular sciences

Article Title: miR-212-5p Regulates PM 2.5 -Induced Apoptosis by Targeting LAMC2 and LAMA3.

doi: 10.3390/ijms26041761

Figure Lengend Snippet: Figure 5. LAMC2 and LAMA3 inhibit apoptosis through the PI3K/AKT/NF-κB pathway: (A) PM2.5- stimulated (180 µg/mL for 24 h) cells transfected with siRNA−LAMC2 or siRNA−LAMA3, and p-IκBα/IκBα and p-p65/p65 protein changes; (B) p-AKT/AKT and p-p85/p85 protein changes; (C) BCL-2, BAD, and caspase-3 protein changes detected with Western blotting; (D) apoptosis rate detected with flow cytometry. The mean ± standard deviation (n = 3) is used to express the data. ns, no significance; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 indicate significance.

Article Snippet: Next, 5% skimmed milk (Beyotime Biotechnology, Shanghai, China) was blocked for 2 h, and the membranes were incubated with specific primary antibodies overnight at 4 ◦C: BAD, BCL-2, caspase-3, LAMC2, p85, AKT, p-AKT, IκBα, β-action (1:1000 dilution, Proteintech, Rosemont, IL, USA), p-IκBα, p65, p-p65 (1:500 dilution, Proteintech, IL, USA), LAMA3, p-p85 (1:500 dilution, Abways, Shanghai, China), and GAPDH (1:7000 dilution, Abways, Shanghai, China).

Techniques: Transfection, Western Blot, Flow Cytometry, Standard Deviation

Journal: Cell

Article Title: Single-cell transcriptomic analysis of primary and metastatic tumor ecosystems in head and neck cancer

doi: 10.1016/j.cell.2017.10.044

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Monoclonal mouse LAMC2, clone CL2980, lot #CL2980 , Novus Biologicals , Cat#NBP2-42388.

Techniques: RNAscope, Labeling, Virus, Recombinant, Cloning, Transfection, Plasmid Preparation, Control, Software

Journal: Journal of Cell Communication and Signaling

Article Title: Restoration of the basement membrane after wounding: a hallmark of young human skin altered with aging

doi: 10.1007/s12079-017-0417-3

Figure Lengend Snippet: Basement membrane protein expression in laser-capture microdissected epidermis and dermis before and during wound healing in young adult human skin. a Unwounded skin baseline levels of COL4A2, COL7A2, and LAMC2 transcripts in epidermis (light gray) and dermis (dark gray) separated by laser capture microdissection. *: p < .05 vs. Epidermis (Wilcoxon signed rank test). b-d Normalized transcript levels in epidermis following wounding (expressed in fold vs. 36B4 (housekeeping gene)) for COL4A2 (b), COL7A2 (c) and LAMC2 (d). N = 3–6 sample per time point. *: p < .05 vs. Epidermis (paired t-test)

Article Snippet: Primer/Probe pairs were obtained from Applied Biosystems (ThermoFisher Scientific) as follows: COL4A2 (Hs01098873_m1), COL7A1 (Hs00164310_m1), and LAMC2 (Hs01043711_m1).

Techniques: Membrane, Expressing, Laser Capture Microdissection

Journal: Journal of Cell Communication and Signaling

Article Title: Restoration of the basement membrane after wounding: a hallmark of young human skin altered with aging

doi: 10.1007/s12079-017-0417-3

Figure Lengend Snippet: Basement membrane protein gene expression in laser-capture microdissected epidermis and dermis of young and aged human skin during wound healing. mRNA levels of LAMC2 (a, d), COL7A2 (b, e), and COL4A2 (c, f) in epidermis (a-c) and dermis (d-f) following wounding (expressed in fold change vs. unwounded baseline). N = 3–6 per group. *: p < .05 vs. Young (unpaired t-test)

Article Snippet: Primer/Probe pairs were obtained from Applied Biosystems (ThermoFisher Scientific) as follows: COL4A2 (Hs01098873_m1), COL7A1 (Hs00164310_m1), and LAMC2 (Hs01043711_m1).

Techniques: Membrane, Gene Expression

Journal: Frontiers in Cell and Developmental Biology

Article Title: Gene Expression Profile of Isolated Dermal Vascular Endothelial Cells in Keloids

doi: 10.3389/fcell.2020.00658

Figure Lengend Snippet: Genes that were highly up- or down-regulated in KVECs compared to in NVECs.

Article Snippet: cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). qRT-PCR assays were performed using an ABI Prism 7500 System (Applied Biosystems, Foster City, CA, United States) with Applied Biosystems TaqMan Gene Expression Assays (SERPINA3, Hs00153674_m1; LAMC2, Hs01043717_m1; GAPDH, Hs03929097_g1) and TaqMan Gene Expression Master Mix (Thermo Fisher Scientific).

Techniques: Sequencing, Membrane

Journal: Frontiers in Cell and Developmental Biology

Article Title: Gene Expression Profile of Isolated Dermal Vascular Endothelial Cells in Keloids

doi: 10.3389/fcell.2020.00658

Figure Lengend Snippet: Network analysis with the 74 and 80 genes that were, respectively, up- and down-regulated by more than 5-fold in keloid vascular endothelial cells. SERPINA3 (red) and LAMC2 (yellow) are located near the center of the network.

Article Snippet: cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). qRT-PCR assays were performed using an ABI Prism 7500 System (Applied Biosystems, Foster City, CA, United States) with Applied Biosystems TaqMan Gene Expression Assays (SERPINA3, Hs00153674_m1; LAMC2, Hs01043717_m1; GAPDH, Hs03929097_g1) and TaqMan Gene Expression Master Mix (Thermo Fisher Scientific).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Gene Expression Profile of Isolated Dermal Vascular Endothelial Cells in Keloids

doi: 10.3389/fcell.2020.00658

Figure Lengend Snippet: Validation of the microarray data by quantitative RT-PCR. The relative mRNA expression levels of SERPINA3 (A) and LAMC2 (B) in keloid vascular endothelial cells (KVECs) ( n = 7) and normal vascular endothelial cells (NVECs) ( n = 6) are shown. All values in this figure represent the mean ± SD.

Article Snippet: cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). qRT-PCR assays were performed using an ABI Prism 7500 System (Applied Biosystems, Foster City, CA, United States) with Applied Biosystems TaqMan Gene Expression Assays (SERPINA3, Hs00153674_m1; LAMC2, Hs01043717_m1; GAPDH, Hs03929097_g1) and TaqMan Gene Expression Master Mix (Thermo Fisher Scientific).

Techniques: Biomarker Discovery, Microarray, Quantitative RT-PCR, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Gene Expression Profile of Isolated Dermal Vascular Endothelial Cells in Keloids

doi: 10.3389/fcell.2020.00658

Figure Lengend Snippet: Immunohistochemical validation of (A) SERPINA3 and (B) LAMC2 protein expression in the blood vessels of keloid and normal skin tissues. Representative histological images are shown. Arrows indicate the blood vessels in the dermis. LPF, low-power field, HPF, high-power field. Scale bars: LPF, 50 μm, HPF, 20 μm.

Article Snippet: cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). qRT-PCR assays were performed using an ABI Prism 7500 System (Applied Biosystems, Foster City, CA, United States) with Applied Biosystems TaqMan Gene Expression Assays (SERPINA3, Hs00153674_m1; LAMC2, Hs01043717_m1; GAPDH, Hs03929097_g1) and TaqMan Gene Expression Master Mix (Thermo Fisher Scientific).

Techniques: Immunohistochemical staining, Biomarker Discovery, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Gene Expression Profile of Isolated Dermal Vascular Endothelial Cells in Keloids

doi: 10.3389/fcell.2020.00658

Figure Lengend Snippet: In vitro study on the biological functions of SERPINA3 and LAMC2 in vascular endothelial cells and fibroblasts. (A) A human dermal vascular endothelial cell line (HMEC-1) was treated with or without 1 μg/ml lipopolysaccharide (LPS) for 15 h ( n = 6). The relative mRNA expression levels of SERPINA3 and LAMC2 in the cells was determined. (B) Normal human dermal fibroblasts (NHDF) were treated with or without 0.5 μg/ml laminin-5 for 15 h ( n = 10). The relative cell proliferation and apoptosis was measured. All values in this figure represent the mean ± SD. ** p < 0.01.

Article Snippet: cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). qRT-PCR assays were performed using an ABI Prism 7500 System (Applied Biosystems, Foster City, CA, United States) with Applied Biosystems TaqMan Gene Expression Assays (SERPINA3, Hs00153674_m1; LAMC2, Hs01043717_m1; GAPDH, Hs03929097_g1) and TaqMan Gene Expression Master Mix (Thermo Fisher Scientific).

Techniques: In Vitro, Expressing

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: LAMC2 in paracancerous and gastric cancer tissues detected by immunohistochemistry. (a) 10 cases of paracancerous tissue samples were collected; the expression of LAMC2 in paracancerous tissue was analyzed by immunohistochemistry; (b) 10 cases of gastric cancer tissue samples were collected; the expression of LAMC2 in gastric cancer tissue was detected through immunohistochemistry.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

Techniques: Immunohistochemistry, Expressing

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: Correlation analysis between the expression of LAMC2, Lnc_AC006548.28, and miR-223 in gastric cancer tissue. (a) The protein level of LAMC2 in different tissues was detected by western blot; (b) the positive percentage of LAMC2 was detected by RT-PCR; it was higher in cancer tissue group than that of paracancerous tissue group; (c) the expression of LAMC2 protein was detected by western blot; the protein level in cancer tissue was higher than that in paracancerous tissue; (d) quantitative PCR was used to detect the expression of LAMC2 mRNA; the expression of Lnc_AC006548.28 in cancer tissues was higher than that in adjacent tissues; (e) the correlation between the expression of LAMC2, Lnc_AC006548.28 and miR-223 was detected by correlation analysis between genes.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

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

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: The size of tumor in different groups. (a) The size of tumor was detected after treatment with lamc2; (b) the tumor growth rate was detected after treatment with lamc2.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

Techniques:

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: Celecoxib exerts antitumor effect through Lnc_ AC006548.28-miR-223-LAMC2 pathway. (a and c) After the treatment with celecoxib or KO Lnc_AC006548.28, OE-miR-223, and KO LAMC2 detected the tumor infiltration by HE staining. The difference was statistically significant. (b and d) Ki67 was used to detect the proliferation of cancer cells; the same results were those that suggest LAMC2 may regulate Lnc_AC006548.28 by regulating miR-223 expression and affect the pathological progress of tumor. (e) Suitable primers were designed. (f) The expression of genes in different groups was detected by PCR test, the difference was statistically significant, and the expression of these genes had the role of mutual regulation.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

Techniques: Staining, Expressing

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: LAMC2 was positively correlated with the expression of Lnc_AC006548.28 and negatively correlated with the expression of miR-223. (a) The suitable cell line and celecoxib concentration were selected by follow-up test. At last, MKN-45 cell and 40 mg/L were used. (b) The cells were collected; the expression of LAMC2 protein and the expression of Lnc_AC006548.28 were detected by western blot. (c) RT-PCR detected the mRNA levels the expression of LAMC2, Lnc_AC006548.28, and miR-223.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

Techniques: Expressing, Concentration Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: Cell proliferation, migration, and invasion detected by CCK8 and Boyden chamber assay. (a) Through inhibiting the expression of LAMC2 and Lnc_AC006548.28 and enhancing the expression of miR-223, cell proliferation was detected by CCK8 experiment; (b, c) through inhibiting the expression of LAMC2 and Lnc_AC006548.28 and enhancing the expression of miR-223, cell migration and invasion ability were detected by Boyden chamber assay. The difference was statistically significant.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

Techniques: Migration, Boyden Chamber Assay, Expressing

Journal: Computational Intelligence and Neuroscience

Article Title: Celecoxib Reverse Invasion and Metastasis of Gastric Cancer through Lnc_AC006548.28-miR-223-LAMC2 Pathway

doi: 10.1155/2022/6140727

Figure Lengend Snippet: Regulatory relationship among Lnc_AC006548.28, miR-223, and LAMC2. (a, b) Through inhibiting the expression of LAMC2 and Lnc_AC006548.28 and enhancing the expression of miR-223, cell apoptosis was detected by flow cytometry experiment; the difference was statistically significant. The regulatory relationship among Lnc_AC006548.28, miR-223, and LAMC2 was detected through Dual-luciferase assay. The difference was statistically significant.

Article Snippet: Primary antibodies against LAMC2 were purchased from Affinity Biosciences, SYBR Premix ExTaq kit (TaKaRa Biotechnology, Dalian, China), TRIzol Reagent (Tiangen, China), and PrimescriptTM RT reagent Kit (TaKaRa Biotechnology, Dalian, China), respectively.

Techniques: Expressing, Flow Cytometry, Luciferase

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A-C) ear scores weekly, from 0 (‘unaffected’) to 6 (‘very affected’). (D-F) tail scores using same method. (G-K) tension at various ages. All mice tested are male, Lamc2 jeb/jeb homozygotes. Congenic names are for Col17a1 ( e.g. , MRL.FVB is MRL- Lamc2 jeb/jeb Col17a1 FVB/FVB ). Ear and tail score statistics are based on age when scores first reach ‘4’ (‘moderately affected’) calculated as survival in PRISM by both Log-rank (Mantel-Cox) and Gehan-Breslow-Wilcoxon, with the less significant of the two indicated. G, MRL required euthanization before 15 wks due to their severity of JEB. Tension statistics are PRISM 1-way ANOVA. Tension bars indicate mean with SD. ns = not significant, * <0.05, ** <0.01, *** <0.001, **** <0.0001. Includes some B6, 129, FVB and MRL data previously published .

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A) Tail tension test at 20 weeks of age. (B-C) Average ear and tail scores from 0 ‘not affected’ to 6 ‘very affected’ for mice scored weekly. (D) Cumulative censored survival data. (E) Amino acid sequence for Col17a1 AA 1275-1292 for tested inbred strains, from Sproule et al 2014. All (A-D) data shown is for FVB males homozygous Lamc2 jeb/jeb and for B6 or MRL Col17a1 congenic segments as indicated. (E) MRL differs from FVB at Col17a1 AA 1277. B6 differs from FVB at both AA 1277 and 1292. Tension bars indicate mean with SD. Tension p-values are 1-way ANOVA. Ear and tail score p-values are less significant of Log-rank (Mantel-Cox) or Gehan-Breslow-Wilcoxon, based on age when each mouse first reaches a score of ‘4’ treated as survival.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques: Sequencing

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A) Tension test results for parental B6 and 129X1, F1 and F2 male mice at 10 weeks of age (P-values: * < 0.05, ** <0.01, **** <0.0001). (B) LOD score of tension x marker analysis for the B6X1F2 tension set. Horizontal lines bottom to top are 63, 10, 5 and 1% cutoffs. (C) Effect plots for SSLP markers nearest the peaks. X-axis is genotype: BB = B6, BX = het, XX = 129X1. (D) 95% confidence intervals for the four most significant QTL. Positions of SSLP markers tested with Mb position names are indicated (i.e. 1_22 is chr1, 22Mb). Blue lines indicate 1.5 LOD drop interval. Green lines indicate Bayesian confidence intervals. 95% CIs converted to Mb are: chr1:22-39Mb (peak 34Mb), chr5:40-111Mb (peak 58Mb), chr11:90-122Mb (peak 119Mb) and chr13:20-104Mb (peak 59Mb). (E) Tail tension comparison of ‘long’ and ‘short’ B6.129X1- Lamc2 jeb/jeb congenics. (F) Mb map of same. 129 is black, B6 is white. All mice are male, Lamc2 jeb/jeb homozygotes. Tension bars indicate mean with SD.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques: Marker

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A) Male 15 week old tail tension test results for MRL.FVB x FVB F 2 (n=176) with F 1 and parental FVB and MRL.FVB controls (all homozygous Lamc2 jeb/jeb and Col17a1 FVB/FVB ). Bars indicate mean with SD. (B) R/QTL LOD scores showing peaks on chr6, 10 and 13. (C) Inclusion of body weight as a covariant increases significance of chr6 and 13 peaks. (D) Effect plots for SSLP markers nearest the QTL peaks. (E) 95% confidence intervals for the three most significant QTL. Converted to Mb are: 6:66-127Mb (peak 110Mb), 10:66-117Mb (peak 104Mb) and 13:11-74Mb (peak 41Mb). Positions of SSLP markers tested with Mb position names are indicated (i.e. 6_26 is chr6, 26Mb). Blue lines indicate 1.5 LOD drop interval. Green lines indicate Bayesian confidence intervals. (F) Linear regression of tension vs body weight for F 2 mice is not statistically significant.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A-F) Mouse Phenome Database (MPD) CGD-MDA1 percent polymorphism plots (PPP) across 1:22-39Mb QTL 95% confidence interval: (A) B6 vs 129X1 percent polymorphic, (B) FVB/MRL and (C) B6/129X1 polymorphic while FVB/MRL matched. Each peak/bar = 0.1 Mb. Map beneath indicates extent of congenic segments tested. Red box indicates Dst position. (G) MPD PPP for Dst . (H) B6 chr1 congenic 10 wk old male tail tension test. All homozygous Lamc2 jeb/jeb . B6.129 and B6.AJ homozygous for chr1 congenic intervals 1:22-37Mb and 1:25-44Mb respectively. Bar indicates mean with SD. (D) Male ear and tail scores for the same congenics. (J) B6/129 missense SNPs mapping to Dst-e . A/J matches B6 at 2 of 5. Statistics in (H) are PRISM 1-way ANOVA all compared to B6- Lamc2 jeb/jeb control (B6) except as indicated. Statistics in (I): both Log-rank (Mantel-Cox) or Gehan-Breslow-Wilcoxon tests were performed, based on age when each mouse first reaches a score of ‘4’ treated as survival. Values shown are the less significant of the two. **** is <0.0001, *** is <0.001, ** is <0.01, * is <0.05, ns is not significant.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A) 10-week-old male tension comparisons of B6, B6- Dst 129/129 congenic, R03Q B6- Col17a1 PWD/PWD congenic, B6- Col17a1 PWD/PWD Dst 129/129 double congenic and B6 wild-type (B6- Lamc2 wt/wt ) (all Lamc2 jeb/jeb except the last). Red symbols indicate clamp slipped off without removing substantial skin. (B) Tension expressed as survival of same with red values censored gives statistical difference between B6- Col17a1 PWD/PWD and double congenic. (C) Photos showing the typical amount of skin removed during the tail tension test from (top to bottom): B6- Lamc2 jeb/jeb , B6- Lamc2 jeb/jeb Dst 129/129 , B6- Lamc2 jeb/jeb Col17a1 PWD/PWD , B6- Lamc2 jeb/jeb Dst 129/129 Col17a1 PWD/PWD double congenic and B6 wild-type, all at 10 wks of age. (D-E) Ear and tail scores for the same excluding B6 wt control and adding FVB- Lamc2 jeb/jeb for comparison. All males. All Lamc2 jeb/jeb except ‘B6- Lamc2 wt/wt ’. Dst is contained within the 1/22-37Mb congenic and used here to represent it. Ear and tail statistics are survival based on age when each mouse first received a score of ‘4’ (moderately affected). **** p<.0001, *** p<.001, ** p<.01, * p<.05, ns = not significant.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A) Percent MPD CGD-MDA1 SNP polymorphism map (each bar = 0.1 Mb) for the chr5 QTL interval. Clustered bars indicate regions of disparate strain heritage. Comparison of B6/129 (top, differ at QTL) to FVB/MRL (middle, predicted to match at QTL) does little to reduce candidate interval (bottom, B6/129 and FVB=MRL). (B) Chr5 congenic 10 wk old male tail tension test. * after name indicates cross made using a reduced B6- Lamc2 jeb/jeb congenic (<10 Mb) instead of originally published (>80 Mb). Statistical significance for each congenic is based on comparison to its parental short or long congenic strain. (C) Focused chr5 congenic map of protective (red) and non-protective (black) lines limits the main candidate interval to 50.5-52.366 Mb. (D) Ensembl view of 5:50.5-52.366Mb showing genes present. (E) Percent polymorphism plots for reduced congenic interval and candidate gene Ppargc1a from the MPD CGD-MDA1 SNP data set.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: (A) Strain heritage comparisons across QTLs: B6 v 129 top, MRL v FVB middle and appropriate prediction-based combined plot bottom for each. Bars indicate percent polymorphism among MPD CGD-MDA1 SNP markers in 0.1Mb increments, only looking at markers which contained genotype information for all 4 strains. Labels indicate full congenic intervals. Red underline indicates location of genes of interest. (B) MPD percent polymorphism for strain pairs for specific candidate intervals and genes. (C) Tail tension test of 10 week old male chr11 QTL congenic vs B6 control. Both groups Lamc2 jeb/jeb . Bars indicate mean with SD.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: bioRxiv

Article Title: Seven naturally variant loci serve as genetic modifiers of Lamc2 jeb induced non-Herlitz Junctional Epidermolysis Bullosa in mice

doi: 10.1101/2023.03.03.530930

Figure Lengend Snippet: Comparison of ear (A, C) and tail (B, D) scores for Col17a1 allele matched B6 and 129X1 versus MRL and FVB matched for Col17a1 FVB/FVB (A, B) or Col17a1 MRL/MRL (C, D). All scores are from 0 (‘unaffected’) to 6 (‘very affected’). (E) Tension comparison of B6, 129, MRL and FVB 10 week old males where B6 and 129 are naturally Col17a1 -matched and MRL and FVB are congenically matched for Col17a1 MRL/MRL . All (A-E) mice are male, homozygous Lamc2 jeb/jeb and homozygous for the indicated alleles of Col17a1 . Ear and tail score statistics (A-D) are less significant of Log-rank (Mantel-Cox) or Gehan-Breslow-Wilcoxon based on age each mouse first to score ‘4’. Tension statistics (E) are 1-way ANOVA. Tension bars indicate mean with SD.

Article Snippet: To reduce the congenic segment and genotyping requirements, B6- Lamc2 jeb/jeb were backcrossed to C57BL/6J two additional generations, selecting for recombinants between Lamc2 and SSLP markers at 150Mb and 160Mb, resulting in a separate stock of B6- Lamc2 jeb/jeb with a <10Mb congenic segment available from The Jackson Laboratory as B6- Lamc2 jeb /DcrJ, JR25467 ( ).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Alcohol promotes epithelial mesenchymal transformation-mediated premetastatic niche formation of colorectal cancer by activating interaction between laminin-γ2 and integrin-β1

doi: 10.3748/wjg.v28.i35.5154

Figure Lengend Snippet: Primer sequences for quantitative real-time polymerase chain reaction

Article Snippet: The following antibodies were used: LAMC2 monoclonal antibody (YT5379, Immunoway, United States), ITGB1 monoclonal antibody (sc-9970, Santa Cruz, United States), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (ab125247, ABCAM, United Kingdom), phosphorylate-FAK (p - FAK)-Y397 (AP0302, ABclonal, Wuhan, China), FAK monoclonal antibody (A11131, ABclonal, Wuhan, China), snail monoclonal antibody (ab53519, ABCAM, United Kingdom), E-cadherin polyclonal antibody (ab1416, ABCAM, United Kingdom), N-cadherin polyclonal antibody (22018-1-AP, Proteintech, United States), fibronectin polyclonal antibody (A16678, ABclonal, Wuhan, China) and special AT-rich sequence binding protein 1 (SATB1) polyclonal antibody (A5800, ABclonal, Wuhan, China).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Alcohol promotes epithelial mesenchymal transformation-mediated premetastatic niche formation of colorectal cancer by activating interaction between laminin-γ2 and integrin-β1

doi: 10.3748/wjg.v28.i35.5154

Figure Lengend Snippet: Alcohol consumption regulates the expression and interaction of integrin-β1 and laminin-γ2 in animal samples. The bars represent the mean ± SD ( n = 8). A and B: Quantification of the protein levels of integrin-β1 (ITGB1) and laminin-γ2 (LAMC2) detected by western blotting in animal samples; C and D: Quantification of the protein levels of ITGB1 and LAMC2 by immunohistochemical (IHC) in animal samples; E: Quantification of the mRNA levels of ITGB1 and LAMC2 by quantitative real-time polymerase chain reaction in clinical samples; F: Statistical analysis of IHC was performed by counting the average optical density (AOD). In Duolink proximity ligation assay (PLA), the brown dot particles observed under the microscope represent the interactions between LAMC2 and ITGB1; G: The statistical analysis of PLA positive signals was performed by counting the AOD under a light microscope. Compared with the non-alcohol group: b P < 0.01, a P < 0.05. ITGB1: Integrin-β1; LAMC2: Laminin-γ2; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; PLA: Proximity ligation assay; IHC: Immunohistochemical staining; AOD: Average optical density.

Article Snippet: The following antibodies were used: LAMC2 monoclonal antibody (YT5379, Immunoway, United States), ITGB1 monoclonal antibody (sc-9970, Santa Cruz, United States), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (ab125247, ABCAM, United Kingdom), phosphorylate-FAK (p - FAK)-Y397 (AP0302, ABclonal, Wuhan, China), FAK monoclonal antibody (A11131, ABclonal, Wuhan, China), snail monoclonal antibody (ab53519, ABCAM, United Kingdom), E-cadherin polyclonal antibody (ab1416, ABCAM, United Kingdom), N-cadherin polyclonal antibody (22018-1-AP, Proteintech, United States), fibronectin polyclonal antibody (A16678, ABclonal, Wuhan, China) and special AT-rich sequence binding protein 1 (SATB1) polyclonal antibody (A5800, ABclonal, Wuhan, China).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Real-time Polymerase Chain Reaction, Proximity Ligation Assay, Microscopy, Light Microscopy, Staining

Journal: World Journal of Gastroenterology

Article Title: Alcohol promotes epithelial mesenchymal transformation-mediated premetastatic niche formation of colorectal cancer by activating interaction between laminin-γ2 and integrin-β1

doi: 10.3748/wjg.v28.i35.5154

Figure Lengend Snippet: Alcohol consumption increased the expression and interaction of laminin-γ2 and integrin-β1 in colorectal cancer rats. The bars represent the mean ± SD ( n = 8). A: Survival analysis of integrin-β1 (ITGB1) and laminin-γ2 (LAMC2) in colorectal cancer; B: The mRNA levels of ITGB1 and LAMC2 determined by quantitative real-time polymerase chain reaction in clinical samples; C: Representative immunohistochemical (IHC) images of ITGB1 and LAMC2 in clinical samples (× 400); D: The quantifications of the protein levels of ITGB1 and LAMC2 determined by IHC in clinical samples; E and F: Duolink proximity ligation assay (PLA) verified the interactions between ITGB1 and LAMC2 in clinical samples. The statistical analysis of PLA positive signals was performed by counting the average optical density under a light microscope. Compared with the non-alcohol group: b P < 0.01, a P < 0.05. LAMC2: Laminin-γ2; ITGB1: Integrin-β1; qRT-PCR: Quantitative real-time polymerase chain reaction; PLA: Proximity ligation assay; IHC: Immunohistochemical staining; AOD: Average optical density. Cancer normal group: cancer adjacent tissue group. NP: No statistical difference.

Article Snippet: The following antibodies were used: LAMC2 monoclonal antibody (YT5379, Immunoway, United States), ITGB1 monoclonal antibody (sc-9970, Santa Cruz, United States), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (ab125247, ABCAM, United Kingdom), phosphorylate-FAK (p - FAK)-Y397 (AP0302, ABclonal, Wuhan, China), FAK monoclonal antibody (A11131, ABclonal, Wuhan, China), snail monoclonal antibody (ab53519, ABCAM, United Kingdom), E-cadherin polyclonal antibody (ab1416, ABCAM, United Kingdom), N-cadherin polyclonal antibody (22018-1-AP, Proteintech, United States), fibronectin polyclonal antibody (A16678, ABclonal, Wuhan, China) and special AT-rich sequence binding protein 1 (SATB1) polyclonal antibody (A5800, ABclonal, Wuhan, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Proximity Ligation Assay, Light Microscopy, Quantitative RT-PCR, Staining

Journal: World Journal of Gastroenterology

Article Title: Alcohol promotes epithelial mesenchymal transformation-mediated premetastatic niche formation of colorectal cancer by activating interaction between laminin-γ2 and integrin-β1

doi: 10.3748/wjg.v28.i35.5154

Figure Lengend Snippet: Schematic diagram of the possible signaling pathway regulated by alcohol. EMT: Epithelial mesenchymal transformation; LAMC2: Laminin-γ2; ITGB1: Integrin-β1; FAK: Focal adhesion kinase; p-FAK: Phosphorylate focal adhesion kinase.

Article Snippet: The following antibodies were used: LAMC2 monoclonal antibody (YT5379, Immunoway, United States), ITGB1 monoclonal antibody (sc-9970, Santa Cruz, United States), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody (ab125247, ABCAM, United Kingdom), phosphorylate-FAK (p - FAK)-Y397 (AP0302, ABclonal, Wuhan, China), FAK monoclonal antibody (A11131, ABclonal, Wuhan, China), snail monoclonal antibody (ab53519, ABCAM, United Kingdom), E-cadherin polyclonal antibody (ab1416, ABCAM, United Kingdom), N-cadherin polyclonal antibody (22018-1-AP, Proteintech, United States), fibronectin polyclonal antibody (A16678, ABclonal, Wuhan, China) and special AT-rich sequence binding protein 1 (SATB1) polyclonal antibody (A5800, ABclonal, Wuhan, China).

Techniques: Transformation Assay