Journal: bioRxiv

Article Title: Disease-linked TDP-43 hyperphosphorylation suppresses TDP-43 condensation and aggregation

doi: 10.1101/2021.04.30.442163

Figure Lengend Snippet: A Immunostainings showing nuclear localization of TDP-43 Wt, 12D and 12A in HeLa cells. Endogenous TDP-43 expression was silenced by siRNAs, followed by transient transfection of the indicated siRNA-resistant myc-TDP-43 constructs. After 24 h, localization of TDP-43 Wt, 12D and 12A variants was visualized by TDP-43 immunostaining (mouse anti-TDP-43 antibody, Proteintech). G3BP1 (rabbit anti-G3BP1 antibody, Proteintech) and DAPI signal is shown to visualize the cytoplasm and nuclei, respectively. In the merge (right column), DAPI is show in turquoise, TDP-43 in green, and G3BP1 in magenta. Bar, 30 μm. B Electrophoretic mobility shift assay (EMSA) of TDP-43-MBP-His 6 variants (Wt, 12D and 12A) in a complex with (UG) 12 RNA. C SDS-PAGE followed by TDP-43 Western blot (upper blot) showing efficient siRNA-mediated knockdown of endogenous TDP-43 (running at ~43 kDa) in comparison to control siRNA and re-expression of myc-tagged TDP-43 Wt, 12D and 12A in Hela cells. Equal loading is demonstrated by α-Tubulin Western blot (bottom). TDP-43 was detected using rabbit anti-TDP-43 C-term antibody (Proteintech), α-Tubulin using mouse anti-alpha Tubulin antibody (Proteintech) and Myc-tag using mouse-anti Myc antibody (9E10, Helmholtz Center Munich).

Article Snippet: Knockdown was analyzed 48 h post transfection by immunohistochemistry using mouse anti TDP-43 antibody (Proteintech, Cat.No: 60019-2-Ig) and immunoblotting using rabbit anti TDP-43 C-Term antibody (Proteintech, Cat.No: 12892-1-AP) to detect TDP-43 and mouse anti alpha-Tubulin antibody (Proteintech, Cat.No: 66031-1-Ig) for detection of α-Tubulin as a control.

Techniques: Expressing, Transfection, Construct, Immunostaining, Electrophoretic Mobility Shift Assay, SDS Page, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance

doi: 10.1155/2019/6471815

Figure Lengend Snippet: Effect of LQZS on MUC5AC synthesis and expression in lung tissues of rats with AECOPD. (a) Immunohistochemical staining of MUC5AC in bronchial epithelium (magnification ×100 and ×400). (b) Estimate of MUC5AC expressions through Western blot. β -actin was used as an internal control. The values were shown as proportions of MUC5AC to β -actin optical density. (c) Changes in relative mRNA levels of MUC5AC. The values were expressed as mean ± SD. One-way ANOVA was adopted for statistical analysis. ∗∗ Compared to Control group P < 0.01; ## compared to AECOPD group P < 0.01; && compared to CAM group P < 0.01.

Article Snippet: Then the filters were incubated with primary antibodies, mice monoclonal IgG anti-rat MUC5AC antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat ROR γ t antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat Foxp3 antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat NE antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat MCP-1 antibody (proteintech, China, dilution 1:1000), and rabbit polyclonal IgG anti-rat β -actin (proteintech, China, dilution 1:2000) served as internal control at 4°C overnight.

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance

doi: 10.1155/2019/6471815

Figure Lengend Snippet: Effect of LQZS on repairing Th17/Treg imbalance in lung tissues of rats with AECOPD. Immunohistochemical staining of ROR γ t (a) and Foxp3 (b) proteins in lung tissues (magnification ×100 and ×400). Estimate of ROR γ t (c) and Foxp3 (d) expressions through Western blot. β -actin was used as an internal control. The values were shown as proportions of target proteins to β -actin optical density. Changes in relative mRNA levels of ROR γ t (e), Foxp3 (f), IL-17(g), IL-10 (h) in lung tissues. The values were expressed as mean ± SD. One-way ANOVA was adopted for statistical analysis. ∗ Compared to Control group P < 0.05, ∗∗ P < 0.01; # compared to AECOPD group P < 0.05, ## P < 0.01; & compared to CAM group P < 0.05, && P < 0.01.

Article Snippet: Then the filters were incubated with primary antibodies, mice monoclonal IgG anti-rat MUC5AC antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat ROR γ t antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat Foxp3 antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat NE antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat MCP-1 antibody (proteintech, China, dilution 1:1000), and rabbit polyclonal IgG anti-rat β -actin (proteintech, China, dilution 1:2000) served as internal control at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Western Blot, Control

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Louqin Zhisou Decoction Inhibits Mucus Hypersecretion for Acute Exacerbation of Chronic Obstructive Pulmonary Disease Rats by Suppressing EGFR-PI3K-AKT Signaling Pathway and Restoring Th17/Treg Balance

doi: 10.1155/2019/6471815

Figure Lengend Snippet: Effect of LQZS on NE and MCP-1 synthesis and expressions in lung tissues of rats with AECOPD. Immunohistochemical staining of NE (a) and MCP-1 (b) proteins in lung tissues (magnification ×100 and ×400). Estimate of NE (c) and MCP-1 (d) expressions through Western blot. β -actin was used as an internal control. The values were shown as proportions of target proteins to β -actin optical density. Changes in relative mRNA levels of NE (e) and MCP-1 (f) in lung tissues. The values were expressed as mean ± SD. One-way ANOVA was adopted for statistical analysis. ∗ Compared to Control group P < 0.05, ∗∗ P < 0.01; # compared to AECOPD group P < 0.05, ## P < 0.01; && compared to CAM group P < 0.01.

Article Snippet: Then the filters were incubated with primary antibodies, mice monoclonal IgG anti-rat MUC5AC antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-EGFR antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-PI3K antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat phospho-AKT antibody (affinity, China, dilution 1:1000), rabbit polyclonal IgG anti-rat ROR γ t antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat Foxp3 antibody (abcam, USA, dilution 1:1000), rabbit polyclonal IgG anti-rat NE antibody (abcam, USA, dilution 1:500), mice monoclonal IgG anti-rat MCP-1 antibody (proteintech, China, dilution 1:1000), and rabbit polyclonal IgG anti-rat β -actin (proteintech, China, dilution 1:2000) served as internal control at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Western Blot, Control