Journal: Melanoma Research

Article Title: Interfering with aggregated α-synuclein in advanced melanoma leads to a major upregulation of MHC class II proteins

doi: 10.1097/CMR.0000000000000982

Figure Lengend Snippet: Proteome and transcriptome changes in human melanoma xenografts treated systemically with anle138b. (a) Comparison of log2FC changes of gene transcript/protein intensities identified by RNA-seq and MS-based proteomics. Candidate transcripts/proteins identified by RNA-seq, MS-based proteomics, or both are colored green, red, and purple, respectively. (b) Correlation of log2FC changes in gene transcript/protein intensity of gene transcripts/proteins selected based upon RNA-seq and MS-based proteomics analysis of WM983-B human melanoma xenografts resected from nude mice that had been treated systemically with anle138b, or had received food pellets not containing anle138b. (c) A tissue section from a nontreated WM983-B melanoma xenograft probed with an antibody to CRYAB. (d) A tissue section from an anle138b-treated WM983-B melanoma xenograft probed with a CRYAB antibody. (e) A tissue section from a nontreated WM983-B melanoma xenograft probed with an antibody to ALDH1A1. (f) A tissue section from an anle138b-treated WM983-B melanoma xenograft probed with an ALDH1A1 antibody. The anti-CRYAB and likewise, the anti-ALDH1A1 antibody-probed tumor xenograft tissue sections (pseudocolored yellow) were counterstained with fluorescent DAPI (pseudocolored blue). (c–f) Scale bar: 50 μm. MS, mass spectrometry.

Article Snippet: Five-μm tissue sections, prepared from cryopreserved WM983-B human melanoma xenografts, were fixed with 4% paraformaldehyde, blocked with 10% goat serum in PBS, and probed with rabbit anti-human antibody against CRYAB (Abcam), ALDH1A1 (Abcam), FN1 (Abcam), or MHC class II (HLA-DPB1) (Abcam).

Techniques: Comparison, RNA Sequencing Assay, Mass Spectrometry

Journal: Stem cell research & therapy

Article Title: Mature human induced pluripotent stem cell-derived cardiomyocytes promote angiogenesis through alpha-B crystallin.

doi: 10.1186/s13287-023-03468-4

Figure Lengend Snippet: Fig. 6 CRYAB was identified as an angiogenic factor of D56-CMs. a Scatter plot of RNA-seq data of D28-CMs and D56-CMs. The red line represents fourfold upregulation in D56-CMs compared with D28-CMs. b qRT-PCR analysis validated the upregulation of CRYAB in D56-CMs. The value for an adult heart sample was set to 1 as a reference. n = 3 per group. c Western blot analysis also confirmed the upregulation of CRYAB protein in D56-CMs compared with D28-CMs. GAPDH protein was used as a loading control. n = 3 per group. d Western blot analysis confirmed the KD of CRYAB (CRYAB-KD) after siRNA treatment in D56-CMs. As a control, non-targeting siRNA was used. n = 3 per group. e CRYAB-KD inhibited HUVEC migration by D56-CMs. The ratio of migrated cells in three independent experiments (n = 3–6 per experiment) is shown. f CRYAB-KD inhibited HUVEC tube lengths by D56-CMs. n = 4 per group. All data are the mean ± SEM, and the P values were determined with unpaired t tests (*P < 0.05, **P < 0.01)

Article Snippet: The Human CRYAB ELISA kit (CUSABIO, Houston, TX, USA; CSB-EL006008HU) was used according to the manufacturer’s protocol.

Techniques: RNA Sequencing, Quantitative RT-PCR, Western Blot, Control, Migration

Journal: Stem cell research & therapy

Article Title: Mature human induced pluripotent stem cell-derived cardiomyocytes promote angiogenesis through alpha-B crystallin.

doi: 10.1186/s13287-023-03468-4

Figure Lengend Snippet: Fig. 7 CRYAB-overexpressing D28-CMs enhanced angiogenesis in vivo. a qRT-PCR analysis validated AAV-mediated CRYAB overexpression (CRYAB-OE) 5 days after infection. An AAV vector carrying only tdTomato (tdTomato-OE) was used as a control. n = 4 per group. The value for an adult heart sample was set to 1 as a reference. b Immunostaining also confirmed significant upregulation of CRYAB in CRYAB-OE grafts (left) compared to tdTomato-OE grafts (right). Grafts are indicated by dotted lines. Scale bars, 100 µm. c Representative images of CD31+ microvessel (green) formation in βMHC+ grafts (red) at 4 weeks post-transplantation. Scale bars, 50 μm. d Quantification of microvessel formation in AAV-infected D28-CM grafts. Five sites were randomly selected from each animal. n = 4 per group. All data are the mean ± SEM, and the P values were determined with unpaired t tests (*P < 0.01, ***P < 0.001)

Article Snippet: The Human CRYAB ELISA kit (CUSABIO, Houston, TX, USA; CSB-EL006008HU) was used according to the manufacturer’s protocol.

Techniques: In Vivo, Quantitative RT-PCR, Over Expression, Infection, Plasmid Preparation, Control, Immunostaining, Transplantation Assay

Journal: Cell Genomics

Article Title: A systems biology approach uncovers novel disease mechanisms in age-related macular degeneration

doi: 10.1016/j.xgen.2023.100302

Figure Lengend Snippet: Pseudo-bulk differential expression from single-nucleus RNA-seq (A–D) Dotplots showing marker gene expression in single-nucleus RNA-seq (sNuc-seq). The color intensity represents the Z score of gene expression, and the dot size represents the percentage of nuclei in each group with non-zero expression. Genes are from pseudo-bulk DE in (A) Müller glia, (B) genes indicating Müller gliosis, (C) RPE, and (D) fibroblasts. (E and F) Examples of pseudo-bulk expression per cell type per donor are in (E) Müller and (F) RPE. (G and H) RNAscope ISH for (G) CRYAB and (H) CLU . Representative images from macular sections of healthy controls (top panels) and GA (middle and bottom panels) from 3 donors each. Nearby sections were stained in (G) and (H), and the images were chosen for close vicinity in each donor. GA images are from the lesion, with lesion centers oriented to the right and the borders to the left. Dashed lines indicate removal of extra white space between the RPE and neural retina that was caused by artifactual postmortem retinal detachment. See also Figure S3 .

Article Snippet: RNAscope probe for human Hs-CRYAB , Advanced Cell Diagnostics , Cat# 426278.

Techniques: Quantitative Proteomics, RNA Sequencing, Marker, Gene Expression, Expressing, RNAscope, Staining

Journal: Cell Genomics

Article Title: A systems biology approach uncovers novel disease mechanisms in age-related macular degeneration

doi: 10.1016/j.xgen.2023.100302

Figure Lengend Snippet:

Article Snippet: RNAscope probe for human Hs-CRYAB , Advanced Cell Diagnostics , Cat# 426278.

Techniques: Recombinant, Protease Inhibitor, RNAscope, DNA Methylation Assay, Sequencing, Methylation, Microscopy, Software