tgfβ1 staining  (ABclonal Biotechnology)

Journal: Nature Communications

Article Title: Nanoparticle-mediated TRPV1 channel blockade amplifies cancer thermo-immunotherapy via heat shock factor 1 modulation

doi: 10.1038/s41467-023-38128-x

Figure Lengend Snippet: CLSM images of HSF1 (green) ( a ) and corresponding co-localization rate analysis of HSF1 with nucleus (blue) ( b ) in PANC02 tumor cells treated with I-Micelles or IS-Micelles under light irradiation (+) or not ( n = 3 independent experiments). Scale bars, 50 μm. CLSM images of TGFβ1 (green) ( c ) and corresponding fluorescence intensity analysis ( d ) of PANC02 tumor sections from the mice treated with different formulations ( n = 3 independent experiments). Scale bars, 50 μm. CLSM images of α-SMA (green) ( e ) and corresponding fluorescence intensity analysis ( f ) of PANC02 tumor sections from the mice treated with different formulations ( n = 3 independent experiments). Scale bars, 50 μm. Masson’s trichrome staining (MTS) ( g ) and corresponding fibrosis analysis ( h ) of PANC02 tumor sections from the mice treated with different formulations ( n = 3 independent experiments). Scale bars, 50 μm. i CLSM images of HSF1 (green) translocation in PANC02 tumor cells treated with IS-Micelles and HSF1A under light irradiation (+), and CLSM images of TGFβ1 (green), α-SMA (green) and Masson’s trichrome staining (MTS) of PANC02 tumor sections from the mice treated with IS-Micelles and HSF1A under light irradiation (+) ( n = 3 independent experiments). Scale bars, 50 μm. CLSM images of Cy5 labeled aPD-L1 (red) ( j ) and corresponding fluorescence intensity analysis ( k ) in PANC02 tumor sections at 24 h post-injection of aPD-L1 from the mice treated with different formulations. Scale bars, 100 μm. This experiment was repeated three times independently with similar results. In c , e , g , and j , the mice were treated with PBS, IS-Micelles, I-Micelles under light irradiation (+), and IS-Micelles under light irradiation (+), respectively. Data are presented as mean ± SD ( b , d , f , h ). Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. Source data are provided as a Source Data file.

Article Snippet: Then, the tissue sections with 10.0 μm thickness were made in a cryostat at 6 h post-irradiation or 24 h post-irradiation or 72 h post-injection, followed by the hematoxylin and eosin (H&E) staining, Ki67 staining (Abcam, ab15580, dilution: 1:100) TUNEL assay (green or red), Masson’s trichrome staining, TGFβ1 staining (Abclonal, A15103, dilution: 1:100), α-SMA staining (Abclonal, A2319, dilution: 1:100), FAPα staining (Abclonal, A6349, dilution: 1:100), collagen I staining (Abclonal, A5786, dilution: 1:100), fibronectin staining (Abclonal, A16678, dilution: 1:100), VEGF-A staining (Abcam, ab52917, dilution: 1:100), and HSP70 staining (Abcam, ab181606, dilution: 1:100) and further labeled with Alexa 594 labeled goat anti-rabbit IgG H&L secondary antibody (Abcam, ab150080, dilution: 1:200) or Alexa 488 labeled goat anti-rabbit IgG H&L secondary antibody (Abcam, ab150077, dilution: 1:200) for 0.5 h in different tissues according to the manufacturer’s protocols.

Techniques: Irradiation, Fluorescence, Staining, Translocation Assay, Labeling, Injection

Journal: Nature Communications

Article Title: Nanoparticle-mediated TRPV1 channel blockade amplifies cancer thermo-immunotherapy via heat shock factor 1 modulation

doi: 10.1038/s41467-023-38128-x

Figure Lengend Snippet: a Timeline for the treatment of subcutaneous PANC02 tumor model. Tumor growth profiles ( b ) and survival curves ( c ) of the mice bearing large PANC02 tumor model (250–300 mm 3 ) treated with various formulations ( n = 7 mice per group). Quantification of CD45 + CD11b + Gr−1 + MDSCs ( d ), CD11b + F4/80 + CD206 + TAMs ( e ) and CD11b + F4/80 + CD80 + TAMs ( f ) inside PANC02 tumors at 72 h post-treatment with various formulations ( n = 5 mice per group). g Ratio of CD11b + F4/80 + CD80 + TAMs to CD11b + F4/80 + CD206 + TAMs inside large PANC02 tumors at 72 h post-treatment with various formulations ( n = 5 mice per group). h Quantification of matured dendritic cells (CD11c + CD80 + CD86 + DCs) inside tumor-draining lymph nodes at 72 h post-treatment with various formulations ( n = 5 mice per group). Quantification of tumor-infiltrating CD45 + CD3 + CD8 + CTLs ( i ) and CD45 + CD3 - CD335 + NK cells ( j ) in PANC02 tumors at 72 h post-treatment with various formulations ( n = 5 mice per group). k Schematic illustration of ECM remodeling to facilitate aPD-L1 infiltration for inducing durable immune response via TRPV1 blockade-synergized thermotherapy. Upon TRPV1 blockade, nuclear translocation of HSF1 is effectively blocked which subsequently suppresses TGFβ1 upregulation and secretion, thus leading to inhibition of CAFs proliferation and activation-mediated ECM proteins deposition, ultimately promoting the infiltration of aPD-L1 and immune cells (e.g. NK cells and T lymphocytes) for durable immune response. Dash lines indicate the failure of downstream signals transduction upon blockade of hyperthermia-activated TRPV1 channel. The mice in b–j were treated with PBS (G1), aPD-L1 (G2), Abraxane/GEM (G3), I-Micelles under light irradiation (+) (G4), I-Micelles/aPD-L1 under light irradiation (+) (G5), IS-Micelles (G6), IS-Micelles/aPD-L1 (G7), IS-Micelles under light irradiation (+) (G8), and IS-Micelles/aPD-L1 under light irradiation (+) (G9), respectively. Data are presented as mean ± SD ( b , d – j ). For b and d – j , statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. For c , statistical significance was calculated by log-rank test. Source data are provided as a Source Data file.

Article Snippet: Then, the tissue sections with 10.0 μm thickness were made in a cryostat at 6 h post-irradiation or 24 h post-irradiation or 72 h post-injection, followed by the hematoxylin and eosin (H&E) staining, Ki67 staining (Abcam, ab15580, dilution: 1:100) TUNEL assay (green or red), Masson’s trichrome staining, TGFβ1 staining (Abclonal, A15103, dilution: 1:100), α-SMA staining (Abclonal, A2319, dilution: 1:100), FAPα staining (Abclonal, A6349, dilution: 1:100), collagen I staining (Abclonal, A5786, dilution: 1:100), fibronectin staining (Abclonal, A16678, dilution: 1:100), VEGF-A staining (Abcam, ab52917, dilution: 1:100), and HSP70 staining (Abcam, ab181606, dilution: 1:100) and further labeled with Alexa 594 labeled goat anti-rabbit IgG H&L secondary antibody (Abcam, ab150080, dilution: 1:200) or Alexa 488 labeled goat anti-rabbit IgG H&L secondary antibody (Abcam, ab150077, dilution: 1:200) for 0.5 h in different tissues according to the manufacturer’s protocols.

Techniques: Translocation Assay, Inhibition, Activation Assay, Transduction, Irradiation

Journal: Development (Cambridge, England)

Article Title: Human embryonic epidermis contains a diverse Langerhans cell precursor pool.

doi: 10.1242/dev.102699

Figure Lengend Snippet: Fig. 4. Skin equivalents fulfil the requirements for LC development. (A) Immunohistochemical staining was performed on cryostat sections of skin equivalents at day 8. TGFβ1 is found throughout the epidermis with occasional cells expressing it at high levels (inset, arrows). Data are representative of five experiments. Scale bar: 50 μm. (B) TGFβ1, GM-CSF and IL-34 levels were analysed in cell culture supernatants of skin equivalents or lysed epidermal equivalents at the indicated time points. Values are means ± s.d. of three experiments. (C) Multiparameter flow cytometry of propagated and cultured CD34+ cord blood-derived haematopoietic stem cells and blood monocytes was performed by incubation with monoclonal antibodies against the cell surface markers indicated (black line, indicated antibody; grey line, isotype control). Gates in dot plots were set according to isotype-matched control staining. Dead cells (based on 7-AAD uptake) were excluded.

Article Snippet: Staining for TGFβ1 (Santa Cruz Biotechnology) and rabbit control serum (Dako) on cryostat section was performed as described previously (Schuster et al., 2009).

Techniques: Immunohistochemical staining, Staining, Expressing, Cell Culture, Flow Cytometry, Derivative Assay, Incubation, Bioprocessing, Control