collagen i  (Cusabio)

Journal: Nature Communications

Article Title: Lipid droplet-associated hydrolase mobilizes stores of liver X receptor sterol ligands and protects against atherosclerosis

doi: 10.1038/s41467-024-50949-y

Figure Lengend Snippet: A qPCR analysis of Abca1 and Abcg1 mRNA in PM isolated from LDAH-Tg (Tg, blue bars) and LDAH-KO (KO, red bars) mice treated with oxLDL (50 μg/ml) for 48 h vs . PM isolated from their respective WT controls (gray bars) (n = 3-4 independent samples). B Common genes identified in RNA-seq analysis of LDAH-KO (red bars) and LDAH-Tg (blue bars) PM vs . their WT controls at q-value < 0.05. C RNA-seq analyzes identified multiple ECM-related genes induced by LDAH. The upper row in the heat map (Tg) represents the ratio of LDAH-Tg PM vs . WT, and the lower row (KO) represents the ratio of LDAH-KO PM vs . their WT controls (n = 4). D qPCR analysis of Col1a1 and Col1a2 genes in LDAH-Tg (Tg, blue bars) and LDAH-KO (KO, red bars) vs . their respective WT control (gray bars) PM treated with oxLDL (50 μg/ml) for 48 h ( n = 3–4). E WT (gray bars) and LDAH-Tg (Tg, blue bars) BMM were treated with siRNA against LXRα (LXR-si) or with non-target siRNA, followed by oxLDL (50 μg/ml) for 48 h. Expression of Nr1h3 (Lxra ), Abca1, Abcg1 , and Col1a1 was determined by qPCR (n = 3). (F) ELISA quantification of pro-collagen I alpha in WT (gray bar) and LDAH-Tg (Tg, blue bar) BMM treated with ox LDL (50 μg/ml) for 48 h ( n = 5–6). G LDAH WT ( gray bars) and KO (red bars) PM were treated with oxLDL (50 μg/mL) for 48 h and media was replaced by media with or without TO901317 (5 μM) for 6 h. TO rescued the downregulation of Abca1, Abcg1 . and Col1a1 seen under LDAH deficiency ( n = 4). H qPCR analysis of Scd1, Fasn and Acaca mRNA in LDAH-Tg (Tg, blue bars) and LDAH-KO (KO, red bars) PM treated with oxLDL (50 μg/ml) for 48 h vs . their respective WT controls (gray bars) ( n = 3-4). In the right panel, WT PM were treated with oxLDL (50 μg/ml) for 48 h and media was replaced by media with ( + TO, purple bars with triangles) or without (-TO, gray bars with circular dots) TO901317 (5 μM) for 6 h (n = 5). Comparisons in ( A ), ( D ), ( E ) and ( H ) were performed by two-tailed unpaired t -test (normally distributed with equal variances), two-tailed Welch’s t -test (normally distributed with unequal variances), or two-tailed Mann-Whitney U (not normally distributed). Comparisons in ( F ) and ( G ) were performed by two-way ANOVA followed by Tukey’s test. *p < 0.05, **p < 0.01, ***p < 0.001. All bars represent mean ± SEM of independent samples. Source data are provided as a Source Data file.

Article Snippet: Fully differentiated BMMs were plated at 2 × 10 6 density on 60 mm dishes and treated with Hi-TBAR oxLDL (Alfa Aesar, J652618PL, 50 μg/ml) for 48 h. Cell lysates were collected using the buffer provided in the Mouse Pro-Collagen I alpha 1 ELISA Kit (Abcam, ab210579).

Techniques: Isolation, RNA Sequencing Assay, Control, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY

Journal: Heliyon

Article Title: Betulin and Crinum asiaticum L. bulbs extract attenuate pulmonary fibrosis by down regulating pro-fibrotic and pro-inflammatory cytokines in bleomycin-induced fibrosis mice model

doi: 10.1016/j.heliyon.2023.e16914

Figure Lengend Snippet: A. Hydroxyproline level in CAE, BET, PFD and BLM treated mice. Data were expressed as mean ± SEM with Dunnett’s multiple comparison test. N = 8, ####ρ < 0.0001 vrs naïve, ****ρ < 0.0001 25, 50 mg/kg BET and 1000 mg/kg CAE vrs BLM. Hydroxyproline – Hyp, Bleomycin induced group – BLM, Crinum asiaticum bulbs extract – CAE, Betulin – BET and Pirfenidone – PFD. B. Collagen I level in CAE, BET, PFD and BLM treated mice. Data were expressed as mean ± SEM with Dunnett’s multiple comparison test. N = 8, ####ρ < 0.0001 for BLM vrs naïve, ****ρ < 0.0001 for 50 mg/kg BET and 1000 mg/kg CAE vrs BLM, ***ρ < 0.005 for 25 mg/kg BET and 500 mg/kg CAE vrs BLM. Bleomycin induced group – BLM, Crinum asiaticum bulbs extract – CAE, Betulin – BET and Pirfenidone – PFD. C. Collagen III level in CAE, BET, PFD and BLM treated mice. Data were expressed as mean ± SEM with Dunnett’s multiple comparison test. N = 8, ####ρ < 0.0001 for BLM vrs naïve, ***ρ < 0.005 for 25, 50 mg/kg BET and 1000 mg/kg CAE vrs BLM, **ρ < 0.05 for 500 mg/kg CAE vrs BLM. Bleomycin induced group – BLM, Crinum asiaticum bulbs extract – CAE, Betulin – BET and Pirfenidone – PFD.

Article Snippet: Bleomycin sulfate (Celon Labs, India, CAT BMI2015AC), Betulin (purity >98% Chem Cruz, USA, CAT: D0522) Pirfenidone (PFD) (Cipla Pharma, India, CAT: P2011), Interleukin-6 ELISA KIT (R and D Systems, UK, CAT: M600B), TNF-alpha ELISA KIT (R and D Systems, UK, CAT: 401-MT), Interleukin-1β (Cusabio, China, CAT: CSB-E08054 m), Hydroxyproline kit (Cusabio, China, CAT: CSB-E08839 m), collagen I and III (Cusabio, China, CAT: CSB-EL005727MO and CSB-E07925 m respectively), TGF-β, MMP-9 (Cusabio, China CAT: CSB-E04726 m), chloroform (BDH Prolabo, CAT: 09H200510).

Techniques: Comparison

Journal: Scientific reports

Article Title: Targeted inhibition of Focal Adhesion Kinase Attenuates Cardiac Fibrosis and Preserves Heart Function in Adverse Cardiac Remodeling.

doi: 10.1038/srep43146

Figure Lengend Snippet: Figure 3. Inhibition of FAK activation decreases ECM synthesis. (a) Western blot evaluated expression of ECM including collagen-1, laminin, fibronectin, as well as LOX. (b) Densitometric analysis of blots for determining ECM normalized to GAPDH. (c) Cell culture medium supernatant was collected and analyzed using a Collagen I alpha 1 ELISA Kit, FAK inhibitor significantly decreased total collagen level in the hypoxia-cultured CFs compared with hypoxia-cultured CFs treated with vehicle only(514.35 ± 63.61 VS. 139.11 ± 48.57 pg/ml). (d) Densitometric analysis of blots for determining LOX to GAPDH. Multiple exposures of LOX are presented in Supplementary Figure S6; full-length blots are presented in Supplementary Figure S3. Data are presented as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: A mouse Collagen I alpha 1 ELISA Kit (Cusabio, Wuhan, China) was used according to the manufacturer’s instructions to determine Collagen I levels.

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay