Journal: Genes to cells : devoted to molecular & cellular mechanisms

Article Title: Src family kinases suppress differentiation of brown adipocytes and browning of white adipocytes.

doi: 10.1111/gtc.12340

Figure Lengend Snippet: Figure 2 Effect of CSK-over-expression on brown adipogenesis. (A–C) Wild-type CSK (CSKWT) and kinase-deficient CSK (CSKK222R) were lentivirally infected to brown preadipocytes 2 days before induction of differentiation. (A) Immunoblotting for CSK (~52 kDa), P-Tyr527-SFK (~55 kDa), and P-Tyr416-SFK (~55 kDa). a-Tubulin (~50 kDa) was blotted as a loading control. (B) Immunostaining for Ucp1 (red). Nuclei were shown by Hoechst staining (blue), and lipid droplets formation was assessed with BODIPY (green). Percentages of Ucp1-positive cells. Scale bars, 100 lm. Values are the means SE (n = 3) *P ≤0.05 vs con- trol, assessed by Student’s t-test. (C) mRNA levels of Ucp1 at days 0, 4, and 7.

Article Snippet: Antibodies were purchased as follows: rabbit anti-Ucp1 (ab10983) antibodies from Abcam; rabbit anti-phospho-Src family (Tyr416) (D49G4) (#6943) and rabbit anti-phospho-Src (Tyr527) (#2105) antibodies from Cell Signaling; rabbit anti-CSK (C20) antibodies from Santa Cruz; mouse anti-a-tubulin (DM1A) antibody from Sigma; Alexa Fluor 594-goat anti-rabbit IgG (#A11037) from Molecular Probes; sheep anti-mouse IgG horseradish peroxidase (HRP)-conjugated (NA9310) and donkey anti-rabbit IgG HRP-conjugated (NA9340) from GE Healthcare.

Techniques: Over Expression, Infection, Western Blot, Control, Immunostaining, Staining

Journal: Oncology reports

Article Title: SRPX and HMCN1 regulate cancer‑associated fibroblasts to promote the invasiveness of ovarian carcinoma.

doi: 10.3892/or.2019.7379

Figure Lengend Snippet: Figure 3. Knockdown of SRPX and HMCN1 in WS1 cells significantly reduced TOV21G cell invasion through the RhoA signaling pathway. Representative multi- ples of changes in comparisons of reverse‑transcription‑quantitative polymerase chain reaction results for (A) SRPX and (B) HMCN1 (***P<0.001). (C) Images of western blot analysis of the expression of SRPX, HMCN1, ROCK, RhoA, cdc42 and MLC in WS1 parental, mock‑transduced, SRPX‑downregulated cells (SRPX‑1 and SRPX‑2) and HMCN1‑downregulated cells (HMCN1‑1 and HMCN1‑2). Similar results were obtained from at least three independent experi- ments. Western blot analysis of HMCN1 was not performed due to no commercially available antibody. SRPX, sushi repeat‑containing protein, X; HMCN1, hemicentin 1; P, parental (normal WS1) cells; Z, mock‑transduced; RhoA, Ras homology family member A; ROCK, Rho kinase; cdc42, cell division cycle 42; MLC, myosin II regulatory light chain; ns, not significant.

Article Snippet: Protein (30 μg) from each well was dissolved in loading sample buffer under reducing conditions, separated by 12% SDS‐PAGE, and transferred to 0.2‐μm nitrocellulose membranes for immunolabelling with the following antibodies: Primary antibodies against SRPX (1:4,000, cat. co. 5473, Cell Signaling Technology, Inc.), RhoA (1:1,000, cat. co. 2117, Cell Signaling Technology, Inc.), ROCK (1:1,000, cat. co. 4035, Cell Signaling Technology, Inc.), cell division cycle 42 (cdc42) (1:1,000, cat. co. 2466, Cell Signaling Technology, Inc.), MLC (1:500, cat. co. 21157, Cell Signaling Technology, Inc.) and β‐actin (1:10,000, cat. co. MA5‐15739, Thermo Fisher Scientific, Inc.) were purchased from Novus Biologicals and incubated at 4 ̊C overnight with β‐actin as a loading control.

Techniques: Knockdown, Polymerase Chain Reaction, Western Blot, Expressing

Journal: Oncology reports

Article Title: SRPX and HMCN1 regulate cancer‑associated fibroblasts to promote the invasiveness of ovarian carcinoma.

doi: 10.3892/or.2019.7379

Figure Lengend Snippet: Figure 4. Overexpression of SRPX in WS1 fibroblasts induces TOV21G cell invasion. Representative (A) changes of SRPX and HMCN1 detected by reverse‑transcription‑quantitative polymerase chain reaction analysis. (B) images of the western blots of the expression of SRPX, ROCK, RhoA and MLC in WS1 parental, mock‑transduced and SRPX‑overexpressing fibroblasts. (C) Overexpression of SRPX in WS1 fibroblasts increased TOV21G cell invasion abili- ties (***P<0.001). The experiment was repeated at least three times. SRPX, sushi repeat‑containing protein, X; HMCN1, hemicentin 1; P, parental (normal WS1) cells; RhoA, Ras homology family member A; ROCK, Rho kinase; cdc42, cell division cycle 42; MLC, myosin II regulatory light chain; ns, not significant.

Article Snippet: Protein (30 μg) from each well was dissolved in loading sample buffer under reducing conditions, separated by 12% SDS‐PAGE, and transferred to 0.2‐μm nitrocellulose membranes for immunolabelling with the following antibodies: Primary antibodies against SRPX (1:4,000, cat. co. 5473, Cell Signaling Technology, Inc.), RhoA (1:1,000, cat. co. 2117, Cell Signaling Technology, Inc.), ROCK (1:1,000, cat. co. 4035, Cell Signaling Technology, Inc.), cell division cycle 42 (cdc42) (1:1,000, cat. co. 2466, Cell Signaling Technology, Inc.), MLC (1:500, cat. co. 21157, Cell Signaling Technology, Inc.) and β‐actin (1:10,000, cat. co. MA5‐15739, Thermo Fisher Scientific, Inc.) were purchased from Novus Biologicals and incubated at 4 ̊C overnight with β‐actin as a loading control.

Techniques: Over Expression, Polymerase Chain Reaction, Western Blot, Expressing