Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: miR-548au-3p promoted rat tracheal chondrocyte proliferation and differentiation. Rat tracheal chondrocytes were transfected with miR-548au-3p mimics, inhibitor, or negative control (NC). (a) Transfection efficiency assessed by PCR. (b) Cell viability of rat tracheal chondrocytes assessed by CCK-8 assay. (c) Cell proliferation ability assessed by EdU staining. Red fluorescence: EdU; blue fluorescence: DAPI (scale bar = 50 μ m). (d) Apoptosis levels assessed by TUNEL assay (scale bar = 50 μ m). Red fluorescence: TUNEL; blue fluorescence: DAPI. (e) Cell apoptosis determined by flow cytometry with Annexin V/propidium iodide staining. (f) Protein levels of E-cadherin and N-cadherin. (g) The effect of miR-548au-3p on differentiation of rat tracheal chondrocytes assessed using Alcian blue staining (scale bar = 100 μ m). (h) Aggrecan, Col2A1, MMP13, and ADAMTS4 protein levels assessed by western blot. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, compared with NC.

Article Snippet: Primary antibodies for this study were specific for CA12 (Boster, Cat. No. A04063, 1 : 1000), LONRF3 (GeneTex, Cat. No. GTX112150, 1 : 2000), MAP2 (Boster, Cat. No. A01201, 1 : 2000), THBS1 (Boster, Cat. No. PB0471, 1 : 2000), PPID, E-cadherin (Beyotime, Cat. No. AF6759, 1 : 1000), N-cadherin (Beyotime, Cat. No. AF5237, 1 : 800), aggrecan (Abcam, Cat. No. ab3778, 1 : 1000), Col2A1 (Boster, Cat. No. A00517, 1 : 2000), MMP13 (Proteintech, Cat. No. 18165-1-AP, 1 : 3000), ADAMTS4 (Proteintech, Cat. No. 11865-1-AP, 1 : 600), and GAPDH (Abcam, Cat. No. ab9485, 1 : 2000).

Techniques: Transfection, Negative Control, CCK-8 Assay, Staining, Fluorescence, TUNEL Assay, Flow Cytometry, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: CA12 suppressed rat tracheal chondrocyte proliferation and differentiation. Rat tracheal chondrocytes were transfected with CA12 overexpression plasmid or CA12 siRNA. (a) Transfection efficiency was determined by western blot. (b) Viability of rat tracheal chondrocytes assessed by CCK-8 assay. (c) Cell proliferation ability assessed by EdU staining. Red fluorescence: EdU; blue fluorescence: DAPI (scale bar = 50 μ m). (d) Cell apoptosis levels assessed by TUNEL staining (scale bar = 50 μ m). Red fluorescence: TUNEL; blue fluorescence: DAPI. (e) Cell apoptosis assessed by flow cytometry with Annexin V/propidium iodide staining. (f) Protein expression of E-cadherin and N-cadherin. (g) The effect of miR-548au-3p on differentiation of rat tracheal chondrocytes assessed with Alcian blue staining (scale bar = 100 μ m). (h) Aggrecan, Col2A1, MMP13, and ADAMTS4 protein levels detected by western blot. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, compared with vector or siRNA NC.

Article Snippet: Primary antibodies for this study were specific for CA12 (Boster, Cat. No. A04063, 1 : 1000), LONRF3 (GeneTex, Cat. No. GTX112150, 1 : 2000), MAP2 (Boster, Cat. No. A01201, 1 : 2000), THBS1 (Boster, Cat. No. PB0471, 1 : 2000), PPID, E-cadherin (Beyotime, Cat. No. AF6759, 1 : 1000), N-cadherin (Beyotime, Cat. No. AF5237, 1 : 800), aggrecan (Abcam, Cat. No. ab3778, 1 : 1000), Col2A1 (Boster, Cat. No. A00517, 1 : 2000), MMP13 (Proteintech, Cat. No. 18165-1-AP, 1 : 3000), ADAMTS4 (Proteintech, Cat. No. 11865-1-AP, 1 : 600), and GAPDH (Abcam, Cat. No. ab9485, 1 : 2000).

Techniques: Transfection, Over Expression, Plasmid Preparation, Western Blot, CCK-8 Assay, Staining, Fluorescence, TUNEL Assay, Flow Cytometry, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: Knockdown of CA12 abolished miR-548au-3p inhibitor mediated effects on rat tracheal chondrocytes. Rat tracheal chondrocytes were cotransfected with CA12 siRNA and miR-548au-3p inhibitor for 48 h. (a) Transfection efficiency was determined by western blot. (b) Viability of rat tracheal chondrocytes assessed by CCK-8 assay. (c) Cell proliferation ability assessed by EdU staining. Red fluorescence: EdU; blue fluorescence: DAPI (scale bar = 50 μ m). (d) Cell apoptosis assessed by TUNEL staining (scale bar = 50 μ m). Red fluorescence: TUNEL; blue fluorescence: DAPI. (e) Cell apoptosis assessed by flow cytometry with Annexin V/propidium iodide staining. (f) E-cadherin and N-cadherin protein levels assessed by western blot. (g) The effect of miR-548au-3p on chondrogenic differentiation of rat tracheal chondrocytes assessed by Alcian blue staining (scale bar = 100 μ m). (h) Aggrecan, Col2A1, MMP13, and ADAMTS4 protein levels assessed by western blot. ∗∗ p < 0.01 and ∗∗∗ p < 0.001, compared with NC or inhibitor siRNA NC.

Article Snippet: Primary antibodies for this study were specific for CA12 (Boster, Cat. No. A04063, 1 : 1000), LONRF3 (GeneTex, Cat. No. GTX112150, 1 : 2000), MAP2 (Boster, Cat. No. A01201, 1 : 2000), THBS1 (Boster, Cat. No. PB0471, 1 : 2000), PPID, E-cadherin (Beyotime, Cat. No. AF6759, 1 : 1000), N-cadherin (Beyotime, Cat. No. AF5237, 1 : 800), aggrecan (Abcam, Cat. No. ab3778, 1 : 1000), Col2A1 (Boster, Cat. No. A00517, 1 : 2000), MMP13 (Proteintech, Cat. No. 18165-1-AP, 1 : 3000), ADAMTS4 (Proteintech, Cat. No. 11865-1-AP, 1 : 600), and GAPDH (Abcam, Cat. No. ab9485, 1 : 2000).

Techniques: Transfection, Western Blot, CCK-8 Assay, Staining, Fluorescence, TUNEL Assay, Flow Cytometry

Journal: Aging Cell

Article Title: Astrocyte Senescence Impairs Synaptogenesis due to Thrombospondin‐1 Loss

doi: 10.1111/acel.70382

Figure Lengend Snippet: The competitive TSP‐1 receptor antagonist gabapentin (GBP) blocks the synaptogenic effect of SAMR1 ACMs. (A and C) Immunostaining of MAP2 (gray), VGLUT1 (red) and PSD95 (green), and quantification of excitatory pre‐ (VGLUT1) and postsynaptic (PSD95) vesicles colocalization in hippocampal neurons treated with ACMs from Diff‐Ast SAMR1 and SAMP8 with or without GBP 32 μM. (B and D) Immunostaining of MAP2 (gray), VGLUT1 (red) and PSD95 (green), and quantification of excitatory pre‐ (VGLUT1) and postsynaptic (PSD95) colocalization vesicles in hippocampal neurons treated with ACMs from ACSA‐2 + SAMR1 and SAMP8 of 6‐m mice with or without GBP 32 μM. Three independent experiments were analyzed per cell type and experimental condition. Data are presented as mean ± SEM. One‐way ANOVA Tukey's multiple comparisons test was performed. * p < 0.05, ** p < 0.01, and *** p < 0.001. Scale bar: 50 μm.

Article Snippet: At 11 DIV, neuron medium was totally replaced by ACM from Ast‐Diff or half replaced by ACSA‐2 + ACM during 3 h. Gabapentin (GBP, MedChemExpress, HY‐A0057) and TSP‐1 (MedChemExpress, HY‐P701325) dosages were based on Cheng et al. ( ) procedures.

Techniques: Immunostaining

Journal: Oncotarget

Article Title: Erbin exerts a protective effect against inflammatory bowel disease by suppressing autophagic cell death.

doi: 10.18632/oncotarget.23925

Figure Lengend Snippet: Figure 5: Expressions of Erbin, LC3, ATG16L1, ATG2A and ATG7 in DSS-induced experimental colitis mice and IL-10 knockout mice (IL-10-/-). (A) Expressions of Erbin (red) and LC3 (green) in colorectum of DSS -treated GFP-LC3+/- mice by immunofluorescent staining and confocal microscopy. The nuclei were stained with DAPI (blue). (B-C) Expressions of Erbin, LC3, ATG16L1, ATG2A and ATG7 by Western Blots (WB) (B), and immunohistochemical staining (IHC) (C). Scale bars = 50 μm (A and C).

Article Snippet: Section and slides were stained for Erbin (A303-762A; Bethyl), LC3B (3868; CST), ATG16L1 (8089; CST), ATG7 (BA3527-2; Boster), ATG2A (15011; CST), respectively.

Techniques: Knock-Out, Staining, Confocal Microscopy, Western Blot, Immunohistochemical staining

Journal: Oncotarget

Article Title: Erbin exerts a protective effect against inflammatory bowel disease by suppressing autophagic cell death.

doi: 10.18632/oncotarget.23925

Figure Lengend Snippet: Figure 6: Autophagic program of colorectum in experiment colitis mouse model after Erbin was deleted. (A) LC3 (green) in colorectum of DSS -treated GFP-LC3+/- mice by immunofluorescent staining and confocal microscopy. The nuclei were stained with DAPI (blue). (B) Ultrastructural changes of autophagic/lysosomal vacuoles (labelled as 1 and mitochondrial (labelled as 2) in colorectal mucosa of Erbin-/- mice treated with DSS. (C-D) Expression of Erbin, LC3, ATG16L1, ATG7, ATG2A of colorectum in DSS-induced wild type (WT) and Erbin-/- mice by Western Blots (WB) (C), and immunohistochemical staining (IHC) in IL-10-/- mice (D). Scale bar, 1 μm and 0.5 μm (B). Scale bar, 50 μm (A and D).

Article Snippet: Section and slides were stained for Erbin (A303-762A; Bethyl), LC3B (3868; CST), ATG16L1 (8089; CST), ATG7 (BA3527-2; Boster), ATG2A (15011; CST), respectively.

Techniques: Staining, Confocal Microscopy, Expressing, Western Blot, Immunohistochemical staining