Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the MMP‐2 levels in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M)+ ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser + E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser+ E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. MMP‐2 levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; MMP‐2, matrix metalloproteinase‐2; P 4 , progesterone.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification

Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the TNF‐α and IL‐6 levels in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M) + ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser + E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser+ E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. (A) represents subsequent time points for TNF‐α analysis, and (B) represents subsequent time points for IL‐6 analysis. TNF‐α and IL‐6 levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; IL‐6: interleukin‐6; MMP‐2, matrix metalloproteinase‐2; P 4 , progesterone; TNF‐α, tumor necrosis factor‐alpha.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification

Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the VEGF‐A levels in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M)+ ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser + E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser+ E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. VEGF‐A levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; P 4 , progesterone; VEGF‐A, vascular endothelial growth factor‐A.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification

Journal: Lasers in Surgery and Medicine

Article Title: Effects of Nonablative Er‐ YAG Laser on Human Endometrial Stromal Cells (hESCs): A Pilot Study

doi: 10.1002/lsm.70020

Figure Lengend Snippet: The boxplots of the IGFBP‐1 secretions in the cultured media of human endometrial stromal cells (hESCs). Confluent hESC cultures were treated with E 2 (10 −8 M) (group E 2 ), E 2 (10 −8 M) + ethanol (0.1%) (group E 2 + S), E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 ), Er‐YAG laser+ E 2 (10 −8 M) (group E 2 + L), Er‐YAG laser + E 2 (10 −8 M) + P 4 (10 −7 M) (group E 2 + P 4 + L) for 12, 24, 48, and 72 h. Only hESCs (incubated in serum‐free DMEM/F12) and hESCs + Er‐YAG laser (hESCs + L) groups were incubated for 12 and 72 h. IGFBP‐1 levels were quantified by ELISA in culture media and normalized to total cell protein ( n = 3, median (Q1–Q3). DMEM, Dulbecco modified Eagle medium; E 2 , estradiol; ELISA, enzyme‐linked immunosorbent assay; Er‐YAG, nonablative Erbium YAG; ESC, endometrial stromal cell; IGFBP‐1, insulin‐like growth factor‐binding protein‐1; P 4 , progesterone.

Article Snippet: To evaluate the remodeling effect of Er‐YAG laser on endometrial tissue, matrix metalloproteinase‐2 (MMP‐2) levels in conditioned media were measured by an enzyme‐linked immunosorbent assay (ELISA) (Elabscience; E‐EL‐H1445, Houston, TX, USA), which has a sensitivity of 0.47 ng/mL and no reported cross‐reactivity or interference [ ].

Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay, Modification, Binding Assay

Journal: Clinical and Translational Medicine

Article Title: Hypoxia‐induced secretory autophagy in cancer‐associated fibroblasts promotes ECM remodelling through serglycin secretion in oral squamous cell carcinoma

doi: 10.1002/ctm2.70556

Figure Lengend Snippet: CAFs secrete SRGN via autophagy to promote OSCC cell invasion and migration by facilitating ECM remodelling through interaction with MMP2/9. (A, B) WB analysis of SRGN protein expression levels and quantification in WT CAFs and SRGN KO CAFs. (C) qPCR analysis of SRGN gene expression in WT CAFs and SRGN KO CAFs. (D) UV image of the agarose gel. (E) The supernatant from normoxic and hypoxic WT CAFs, WT CAFs + 3‐MA, and SRGN KO CAFs was collected and co‐cultured with OSCC cells. Invasion ability was assessed by transwell assays. (F) Invasion cell numbers were quantified using ImageJ software. (* p < .05; ** p < .01; *** p < .001). (G) The supernatant from normoxic and hypoxic WT CAFs, WT CAFs + 3‐MA, and SRGN KO CAFs was collected and co‐cultured with SCC9 cells. Migration ability was assessed by scratch assays. (H) Prediction of SRGN‐binding proteins using the STRING database. (I) HEK293T cells were transfected with SRGN‐Flag and incubated for 48 h. Cell lysates were incubated with anti‐Flag beads, and immunoblotting (IB) was performed using anti‐Flag, anti‐MMP11, anti‐MMP9, and anti‐MMP2 antibodies. (J) WB analysis of changes in MMP9, MMP11, MMP2, and SRGN protein expression levels in WT CAFs and SRGN KO CAFs. (K) Gelatin degradation assays were performed to evaluate gelatin degradation after 24 h of co‐culture of CAL27 cells with the supernatants from normoxic and hypoxic WT CAFs, WT CAFs + 3‐MA, and SRGN KO CAFs. Scale bar = 20 µm.

Article Snippet: Primary antibodies used in this assay included: β‐actin (1:4000, 20536‐1, Proteintech), MMP9 (1:1000, bs‐4593R, Bioss), MMP2 (1:1000, CY5189, Abways), MMP11 (1:1000, CY5778, Abways), Flag (1:5000, 80801‐2‐RR, Proteintech).

Techniques: Migration, Expressing, Gene Expression, Agarose Gel Electrophoresis, Cell Culture, Software, Binding Assay, Transfection, Incubation, Western Blot, Co-Culture Assay

Journal: Clinical and Translational Medicine

Article Title: Hypoxia‐induced secretory autophagy in cancer‐associated fibroblasts promotes ECM remodelling through serglycin secretion in oral squamous cell carcinoma

doi: 10.1002/ctm2.70556

Figure Lengend Snippet: CAF‐derived SRGN promotes tumour invasion and ECM degradation via autophagy secretion. (A) In vivo xenograft models were established in nude mice and divided into four groups: (a) CAL27, (b) CAL27 + WT CAFs, (c) CAL27 + WT CAFs (3‐MA), and (d) CAL27 + SRGN KO CAFs. (B) Tumour volume and tumour weight were monitored ( n = 7). (C, D) H&E staining and IHC analysis of COL1, E‐cadherin, MMP2 and MMP9 were performed in orthotopic xenograft tumour tissues. The expression levels of COL1, E‐cadherin, MMP2 and MMP9 were quantitatively analyzed using Fiji software. Scale bar = 100 µm. (* p < .05; ** p < .01; *** p < .001).

Article Snippet: Primary antibodies used in this assay included: β‐actin (1:4000, 20536‐1, Proteintech), MMP9 (1:1000, bs‐4593R, Bioss), MMP2 (1:1000, CY5189, Abways), MMP11 (1:1000, CY5778, Abways), Flag (1:5000, 80801‐2‐RR, Proteintech).

Techniques: Derivative Assay, In Vivo, Staining, Expressing, Software

Journal: Clinical and Translational Medicine

Article Title: Hypoxia‐induced secretory autophagy in cancer‐associated fibroblasts promotes ECM remodelling through serglycin secretion in oral squamous cell carcinoma

doi: 10.1002/ctm2.70556

Figure Lengend Snippet: Mechanism diagram of hypoxic CAFs‐derived SRGN secretion and tumour progression promotion. Under normal conditions, SRGN is translocated into the ER and subsequently transported via the Golgi apparatus for secretion into the extracellular space. Under hypoxic conditions, elevated autophagy levels in CAFs facilitate the release of SRGN into the ECM through secretory autophagy‐mediated plasma membrane fusion. Within the ECM, SRGN interacts with MMP2 and MMP9, enhancing ECM remodelling and ultimately promoting the invasive capacity of OSCC cells.

Article Snippet: Primary antibodies used in this assay included: β‐actin (1:4000, 20536‐1, Proteintech), MMP9 (1:1000, bs‐4593R, Bioss), MMP2 (1:1000, CY5189, Abways), MMP11 (1:1000, CY5778, Abways), Flag (1:5000, 80801‐2‐RR, Proteintech).

Techniques: Derivative Assay, Clinical Proteomics, Membrane

Journal: ACS nano

Article Title: Echinacoside-Zinc Nanomaterial Inhibits Skin Glycation by Suppressing the Transcriptional Activation of the Receptor for Advanced Glycation End-Products.

doi: 10.1021/acsnano.3c04726

Figure Lengend Snippet: Figure 2. PPZn exerted anti-antiglycan in glycated model mouse skin. (A−C) Representative images of immunohistochemical (IHC) staining (A−a), HE staining (B−a), and Masson staining (C−a) of mouse skin tissue after the indicated treatment. The statistical results of the IHC staining index (A−b), epidermal thickness (B−b), and collagen density (C−b) are shown on the right. Scale bar, 50 μm. (D) qRT- PCR detection of RAGE mRNA levels in skin tissues of mice after the indicated treatment. (E) Western blot representative images and quantitative analysis of RAGE, COL1A2, MMP1, AGEs, and β-actin protein levels in skin tissues of mice after the indicated treatment. (F) Representative images of TUNEL staining of mouse skin tissue after the indicated treatment. Scale bar, 50 μm. (G−I) Relative Hyp content (G), SOD activity (H), and MDA concentration (I) in skin tissues of mice after the indicated treatment. All values are presented as the mean ± SD; P-values determined by two-sided Student’s t test. *P < 0.05, **P < 0.01, compared with the model.

Article Snippet: After being blocked, samples were then incubated with primary antibodies against MDM2 (Proteintech, 1:200), RAGE (Santa, 1:50), STAT2 (Zenbio, 1:100), MMP1 (Affinity, 1:100), COL1A2 (Proteintech, 1:100), and AGEs (Abcam, 1:100) at 4 °C overnight.

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Quantitative RT-PCR, Western Blot, TUNEL Assay, Activity Assay, Concentration Assay

Journal: ACS nano

Article Title: Echinacoside-Zinc Nanomaterial Inhibits Skin Glycation by Suppressing the Transcriptional Activation of the Receptor for Advanced Glycation End-Products.

doi: 10.1021/acsnano.3c04726

Figure Lengend Snippet: Figure 3. PPZn exerted antiglycation effects in HaCaT cells. (A) Western blot representative images and quantitative analysis of RAGE, COL1A2, MMP1, AGEs, and β-actin protein levels in HaCaT cells after the indicated treatment. (B) Cell cycle was determined by flow cytometry in HaCaT cells after the indicated treatment. (C) Representative images of TUNEL staining of HaCaT cells after the indicated treatment. Scale bar, 50 μm. (D, E) Relative MDA concentration (D) and SOD activity (E) in HaCaT cells after the indicated treatment. All values are presented as the mean ± SD; P-values determined by two-sided Student’s t test. *P < 0.05, **P < 0.01, compared with the model.

Article Snippet: After being blocked, samples were then incubated with primary antibodies against MDM2 (Proteintech, 1:200), RAGE (Santa, 1:50), STAT2 (Zenbio, 1:100), MMP1 (Affinity, 1:100), COL1A2 (Proteintech, 1:100), and AGEs (Abcam, 1:100) at 4 °C overnight.

Techniques: Western Blot, Flow Cytometry, TUNEL Assay, Staining, Concentration Assay, Activity Assay

Journal: Cell Proliferation

Article Title: Soluble Sema4D From γδ T Cells Exerts Osteoblast Inhibition via Plexin‐B/ mTOR Signalling Contributing to Pathogenesis of Bisphosphonate‐Related Osteonecrosis of the Jaws

doi: 10.1111/cpr.70114

Figure Lengend Snippet: Autocrine MMP3 secretion induced mSema4D cleavage in γδ T cells. (A) KEGG analysis of DEGs in WT ZOL and TCRδ −/− ZOL mice was highly enriched in matrix metalloproteinase (MMP)‐related pathways. (B) The mRNA levels of Mmp3 and Adam17 in the extraction sockets of WT ZOL and TCRδ −/− ZOL mice. n = 4, * p < 0.05, ns indicates no significance. (C) The mRNA levels of Adam17 , Adam10 , Mmp2 , Mmp3 , Mmp9 and Mmp11 after γδ T cells were incubated with 15 μM ZOL for 3 and 6 days. Only the mRNA level of Mmp3 increased significantly and kept a high level after 6 days incubation. n = 6, * p < 0.05, ** p < 0.01. (D) The protein level of MMP3 was much higher than MMP2 and MMP9 in the γδ T cell supernatant measured by ELISA. n = 4, ** p < 0.01, ns indicates no significance. (E) The proportions of γδ T cells after treatment with GM6001 and TAPI‐2. GM6001 and TAPI‐2 are MMP inhibitors. T10 and T20 indicate 10 and 20 nM TAPI‐2, respectively; G50 and G100 indicate 50 and 100 nM GM6001, respectively. n = 6, ns indicates no significance. (F) The expression of Sema4D on γδ T cells after treatment with GM6001 and TAPI‐2. (G) Analysis of Sema4D expression in γδ T cells after treatment with GM6001 or TAPI‐2 on day 2 and 4. n = 6, * p < 0.05, ** p < 0.01. (H) The level of sSema4D in the supernatant of γδ T cells after treatment with GM6001 or TAPI‐2. n = 3, * p < 0.05, ** p < 0.01. (I) The level of sSema4D in the supernatant of γδ T cells after incubating with MMP2, MMP3 and MMP9 in synergy with GM6001. n = 4, * p < 0.05, ** p < 0.01. Data are mean ± SD.

Article Snippet: The kits of sSema4D (ELK5378, ELK Biotechnology), MMP2 (E‐EL‐M0780, Elabscience), MMP3 (E‐EL‐M0626) and MMP9 (E‐EL‐M3052) were used following the manufacturer's instructions.

Techniques: Extraction, Incubation, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Cell reports

Article Title: CRISPR activation screens identify the SWI/SNF ATPases as suppressors of ferroptosis.

doi: 10.1016/j.celrep.2024.114345

Figure Lengend Snippet: Figure 4. Ferroptosis resistance and NRF2 activation are conserved functions between BRM and BRG1 and require their ATPase activity (A) Colony-forming assays of KP4 cells overexpressing WT BRG1, catalytic mutants K785R or T910M BRG, or LACZ. Scale bar, 1 mm. Representative of n = 3 biological replicates is depicted. (B) BODIPY-C11 imaging of lipid peroxidation levels in the indicated cells. Each dot represents a cell, and the red lines represent the median. p values were calculated by a Kruskal-Wallis ANOVA with a Dunn’s posttest. Experiments in (A) and (B) were repeated at least 3 times with similar results. (C) Pile-up plots of ATAC-seq-accessible peaks in KP4 cells expressing WT BRG1 versus KR BRG1 mutant, TM BRG1 mutant, or LACZ (n = 2 biological rep- licates). See also Figure S4E and Table S6.

Article Snippet: For cut&run, the following antibodies were used – BRM (Epicypher, 13–2006), NRF2 (Active Motif, AB_2793695), SMARCC1 (Cell Signaling, 11956), H3K4me1 (Epicypher, 13–0057), H3K27ac (Cell Signaling, 8173), IgG (Epicypher, 13–0042) and H3K4me3 (Epicypher, 13–0041) at the recommended concentrations.

Techniques: Activation Assay, Activity Assay, Imaging, Expressing, Mutagenesis

Journal: Cell reports

Article Title: CRISPR activation screens identify the SWI/SNF ATPases as suppressors of ferroptosis.

doi: 10.1016/j.celrep.2024.114345

Figure Lengend Snippet: Figure 5. The SWI/SNF complex binds to NRF2 sites at enhancers and promoters to increase chromatin accessibility and gene expression (A and G) Pile-up plots of normalized mean read counts for the indicated antibodies from Cut&Run experiments (n = 2 biological replicates) generated using consensus sites. (B, C, H, I) Venn diagram of gene-level overlap between the indicated factors. The number of regions are indicated for each condition. For (B), p = 1e1,037 for overlap between NRF2 and BRM, p = 1e1,291 for overlap between BRM, NRF2, and SMARCC1, and p = 1e2,064 for overlap between BRM and SMARCC1. For (H), p = 1e504 for overlap between NRF2 and BRG1, p = 1e145 for overlap between BRG1, NRF2, and SMARCC1, and p = 1e135 for overlap between BRG1 and SMARCC1. For (B) and (H), p values were determined using hypergeometric distribution. (D and J) Genome browser view of BRM Cut&Run (D) or BRG1 Cut&Run (J) for the indicated antibodies at TKT, a NRF2 target gene.

Article Snippet: For cut&run, the following antibodies were used – BRM (Epicypher, 13–2006), NRF2 (Active Motif, AB_2793695), SMARCC1 (Cell Signaling, 11956), H3K4me1 (Epicypher, 13–0057), H3K27ac (Cell Signaling, 8173), IgG (Epicypher, 13–0042) and H3K4me3 (Epicypher, 13–0041) at the recommended concentrations.

Techniques: Gene Expression, Generated

Journal: Cell reports

Article Title: CRISPR activation screens identify the SWI/SNF ATPases as suppressors of ferroptosis.

doi: 10.1016/j.celrep.2024.114345

Figure Lengend Snippet: Figure 6. The SWI/SNF complex enhances NRF2 binding to the chromatin (A) Representative western blot showing NRF2, TUBULIN, Histone H3, and BRG1 protein in the indicated cellular fractions in KP4 cells. (B) Bar graph quantifications (mean ± SEM, n = 3 biological replicates) showing relative chromatin to cytoplasmic ratio of NRF2 protein from 3 independent experiments. The chromatin to cytoplasmic ratio of NRF2 in LACZ-expressing cells was set to 1. p values were determined by a 1-way ANOVA test with Tukey’s correction for multiple comparisons. (C) Representative ML210 dose-response curves (mean ± SEM, n = 3 independently treated wells) of cells overexpressing BRM transduced with a sgRNA targeting NRF2 (sgNRF2) or a non-targeting sgRNA (sgNT). p values derived from a 2-way ANOVA with Tukey’s correction for multiple comparisons.

Article Snippet: For cut&run, the following antibodies were used – BRM (Epicypher, 13–2006), NRF2 (Active Motif, AB_2793695), SMARCC1 (Cell Signaling, 11956), H3K4me1 (Epicypher, 13–0057), H3K27ac (Cell Signaling, 8173), IgG (Epicypher, 13–0042) and H3K4me3 (Epicypher, 13–0041) at the recommended concentrations.

Techniques: Binding Assay, Western Blot, Expressing, Transduction, Derivative Assay