Journal: Journal of nanobiotechnology

Article Title: Evaluating the pro-survival potential of apoptotic bodies derived from 2D- and 3D- cultured adipose stem cells in ischaemic flaps.

doi: 10.1186/s12951-024-02533-1

Figure Lengend Snippet: Fig. 6 3D-ABs suppress oxidative stress and cell death, and stimulated angiogenesis in ischaemic flaps. (A) F-CHP staining for damaged collagen detec tion in the skin on POD7. Scale bars, 100 μm. (B) The three groups’ quantified F-CHP intensity (n = 6). (C) CD31 and α-SMA IF staining in the flap on POD7. Scale bars, 50 μm. (D) Quantified CD31/α-SMA-positive blood vessel density among the three groups (n = 6). (E) Dead cells in flap tissue sections on POD7 under the detection of TUNEL staining. Scale bar, 20 μm. (F) TUNEL-positive cell percentage in the dermal layer, quantified for each of the three groups (n = 6). (G) DHE staining of the skin tissues on POD7 among the three groups. Scale bars, 50 μm. (H) Quantified DHE among the three groups (n = 6). SEM error bars are used. Significance (*): p value < 0.05; equal variances ANOVA with LSD post hoc analysis or unequal variances Dunnett’s T3 technique

Article Snippet: The primary antibodies utilized were specific to CD31 (1:200), CD68 (1:200), α-SMA (1:200; Proteintech, 67735-1-Ig), iNOS (1:200), and Arg1 (1:200).

Techniques: Staining, TUNEL Assay

Journal: Cells

Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

doi: 10.3390/cells13232005

Figure Lengend Snippet: TGFβ induces WISP1 expression in primary lung and dermal fibroblasts. Stimulation with TGFβ (1 ng/mL) for 24 h significantly increases COL1A1, COL3A1, IL6, ACTA2, FN1, and WISP1 mRNA as compared to control in ( A ) NHLF, ( B ) DHLF, and ( C ) NHDF for 3 respective donors ( n = 3/each) analyzed by RT-qPCR. Notably, TGFβ decreased CCL2 in NHLF and DHLF, except in NHDF. Each experiment was independently performed in triplicates with three biological replicates ( n = 3) for each donor and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the unstimulated control, as shown.

Article Snippet: The following commercially available TaqMan probes and primers were utilized: human Col1A1 (Hs00164004_m1), Col3A1 (Hs00943809_m1), CCL2 (Hs00234140_m1), IL6 (Hs00985639_m1), ACTA2 (Hs05005341_m1), FN1 (Hs01549976_m1), CCN4/WISP1 (Hs00987448_m1), and 18S (4310893E) from Applied Biosystems. cDNAs (1:20 dilution ratio) were added along with TaqMan Fast-advanced Mastermix in a total 10 μL reaction volume in a 384-well plate as per the manufacturer’s instructions and the plate was read using Quant Studio 7 Flex (Applied Biosystems, Waltham, MA, USA).

Techniques: Expressing, Control, Quantitative RT-PCR

Journal: Cells

Article Title: Stage-Dependent Fibrotic Gene Profiling of WISP1-Mediated Fibrogenesis in Human Fibroblasts

doi: 10.3390/cells13232005

Figure Lengend Snippet: WISP1 increases CCL2 and IL6 in primary fibroblasts. Stimulation with WISP1 (1000 nM) for 24 h significantly increases CCL2 and IL6 gene expression in ( A ) NHLF, ( B ) DHLF, and ( C ) NHDF as compared to control. TGFβ (1 ng/mL) for 24 h served as a positive control and significantly increases COL1A1, COL3A1, IL6, ACTA2, FN1, and WISP1 versus vehicle control. ( D ) Stimulation with increasing WISP1 concentrations for 24 h initiates concentration-dependent increase in CCL2 and IL6 protein levels, detected via ELISA in the cell-supernatant of NHDF. Each experiment performed in duplicates with three biological replicates ( n = 3) and represented as mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc analysis vs. vehicle control for respective donors. * denotes p < 0.5, ** denotes p < 0.01, *** denotes p < 0.001, **** denotes p < 0.0001 versus the vehicle control, as shown.

Article Snippet: The following commercially available TaqMan probes and primers were utilized: human Col1A1 (Hs00164004_m1), Col3A1 (Hs00943809_m1), CCL2 (Hs00234140_m1), IL6 (Hs00985639_m1), ACTA2 (Hs05005341_m1), FN1 (Hs01549976_m1), CCN4/WISP1 (Hs00987448_m1), and 18S (4310893E) from Applied Biosystems. cDNAs (1:20 dilution ratio) were added along with TaqMan Fast-advanced Mastermix in a total 10 μL reaction volume in a 384-well plate as per the manufacturer’s instructions and the plate was read using Quant Studio 7 Flex (Applied Biosystems, Waltham, MA, USA).

Techniques: Gene Expression, Control, Positive Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Acta biomaterialia

Article Title: Tumor necrosis factor alpha and interleukin 1 beta suppress myofibroblast activation via nuclear factor kappa B signaling in 3D-cultured mitral valve interstitial cells

doi: 10.1016/j.actbio.2021.03.075

Figure Lengend Snippet: TNF-α and IL-1β decrease activation-related gene expression in 3D-cultured mitral VICs. (A) RNA expression of ACTA2, which encodes the activated VIC marker αSMA, in 3D-cultured mitral VICs treated with 10 ng/ml TNF-α or IL-1β for 2 days in the presence or absence of 40 μM PS-1145 (“PS”). (B) RNA expression of TAGLN, which encodes the activated VIC marker SM22α, under the same treatment conditions. ∗ p < 0.05 (n = 4 samples per group). Relative expression levels are normalized to GAPDH.

Article Snippet: Quantitative reverse transcriptase PCR was performed using the iTaq Universal Probes Supermix (Bio-Rad, Hercules, CA) with TaqMan primers specific to porcine ACTA2 (probe ID: Ss04245588_m1), TAGLN (Ss03373216_g1), TGFB1 (Ss04955543_m1), COL1A1 (Ss03373340_m1), COL3A1 (Ss04323794_m1), and GAPDH (Ss03374854_g1).

Techniques: Activation Assay, Expressing, Cell Culture, RNA Expression, Marker