Journal: International Journal of Molecular Sciences

Article Title: Widespread Expression of Hedgehog Pathway Components in a Large Panel of Human Tumor Cells and Inhibition of Tumor Growth by GANT61: Implications for Cancer Therapy

doi: 10.3390/ijms19092682

Figure Lengend Snippet: Panel of protein expression pattern of HH signaling components. Western blots made in RIPA extracts (30 μg) were probed with indicated antibodies. With some small exceptions, all HH proteins were expressed, although sometimes the expression level was weaker (see text). Survivin, an HH target, was invariably present in tumor cell lines. Notably, GLI3 was shown as a fragment that was cleaved off from the whole protein during sample preparation. However, its signals represent the true amount of intact GLI3 in the extract. The size of each protein in shown in kDa on the right.

Article Snippet: Commercially available primary antibodies used were as follows: Sonic Hedgehog, Cell Signaling Technology #2207 (Danvers, MA, USA); Patched, Biorbyt #157169 (San Francisco, CA, USA); SMO, #ab72130 (Abcam, Cambridge, UK); SuFu, Cell Signaling #2520; Gli, Abcam #ab134906; Gli2, #sc-271786 (Santa Cruz Biotechnology, Dallas, TX, USA); Gli3, Biorbyt #157158; Survivin, Santa Cruz #sc-17779; BCL-2, BD Pharmingen #556354 (San Jose, CA, USA); β-actin, Sigma #A5316.

Techniques: Expressing, Western Blot, Sample Prep

Journal: Animals : an Open Access Journal from MDPI

Article Title: Myogenic Determination and Differentiation of Chicken Bone Marrow-Derived Mesenchymal Stem Cells under Different Inductive Agents

doi: 10.3390/ani12121531

Figure Lengend Snippet: Induction of myogenic differentiation in chicken BM-MSCs under different inductive agents. Muscle-related genes were detected using qRT-PCR after ( a ) DXMS and HCor and ( b ) 5-Aza treatment for 1 day, 3 days, 5 days, or 7 days. ( c ) Based on the qRT-PCR data, the 5-Aza-treated group showed a better myogenic effect in comparison with the DXMS-HC-treated group. ( d ) Western blot results showed that MyoD1 and MyoG were significantly upregulated with the treatment of 5-Aza for 5 days. ( e ) Myogenic protein expression assessed by Western blot analysis is expressed relative amount to Tubulin. Symbol “*” and “**” indicate a significant difference at p < 0.05 and p < 0.01, respectively.

Article Snippet: The following antibodies and their dilutions were used in the Western blot: rabbit anti-PPAR gamma polyclonal antibody (bs0530R, Bioss, Zhuhai, China, 1:1000); rabbit anti-CEBP alpha polyclonal antibody (bs-24540R, Bioss, Zhuhai, China, 1:1000); rabbit anti-beta-actin polyclonal antibody (bs-0061R, Bioss, Zhuhai, China, 1:5000); MyoD1 monoclonal antibody (PM2147a, GenePharma, Suzhou, China, 1:100); MyoG polyclonal antibody (orb6492, Biorbyt, Cambridge, UK, 1:500); HRP, Goat Anti-Rabbit IgG polyclonal (A21020, Abbkine, Wuhan, China, 1:10,000); and HRP, Goat Anti-Mouse IgG polyclonal (A21010, Abbkine, Wuhan, China, 1:10,000).

Techniques: Quantitative RT-PCR, Western Blot, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Hyperphosphorylation of ribosomal protein S6 predicts unfavorable clinical survival in non-small cell lung cancer

doi: 10.1186/s13046-015-0239-1

Figure Lengend Snippet: The upstream regulation signaling pathway of rpS6. a Separate overexpression of Akt1, Akt2 and Akt3 in HBE cells showed that only Akt2 (oe-Akt2) led to marked hyperphosphorylation of mTOR, p70S6K and rpS6 (left; All P < 0.05); while Akt1 or Akt3 alteration resulted in no detectable change of these proteins (left; All P > 0.05). In H1650 and SK-MES-1 cell lines, only the specific silence of Akt2 (sh-Akt2) caused significant loss of p-mTOR, p-p70S6K and p-rpS6 (middle and right; All P < 0.05), and no notable alteration with the Akt1 and Akt3 knockdown were found (sh-Akt1, sh-Akt3; middle and right; All P < 0.05). b H1650 and SK-MES-1 were treated with Akti-2 (Akt inhibitor XII) for 72 h in concentration of 0.8, 3.2 and 12.8 μM. Western blot assays revealed the strongest effect of 12.8 μM and the accompanied dephosphorylation of mTOR, p70S6K and rpS6 (All P < 0.05). oe-Akt1: overexpression of Akt1; oe-NCa1: negative control for oe-Akt1; oe-Akt2: overexpression of oe-Akt2; oe-NCa2: negative control for oe-Akt2; oe-Akt3: overexpression of oe-Akt3; oe-NCa3: negative control for oe-Akt3. sh-Akt1: knockdown for Akt1; sh-NCa1: negative control for sh-Akt1; sh-Akt2: knockdown for Akt2; sh-NCa2: negative control for sh-Akt2; sh-Akt3: knockdown for Akt3; sh-NCa3: negative control for sh-Akt3

Article Snippet: Specific protein antibodies included anti-total rpS6 (CST, #2217), anti-p-rpS6 (Ser235/236, CST, #4858), anti-p21 Cip1 (CST, #2947), anti-p27 Kip1 (CST, #3688), anti-CDK2 (CST, #2546), anti-CDK4 (CST, #12790), anti-Cyclin E (SAB, #29030), anti-cyclin D1 (CST, #2978), anti-p-Rb (Ser780, CST, #3590), anti-Paxillin (CST, #12065), anti-p-Paxillin (Tyr118; CST, #2541), anti-vimentin (CST, #5741), anti-N-cadherin (CST, #4061), anti-E-cadherin (CST, #3195), anti-MMP-9 (CST, #13667), anti-MMP-2 (CST, # 5741), anti-t-Akt1 (CST, #2967), anti-p-Akt1 (Ser473, CST, # 9018), anti-t-Akt2 (CST, #2964), anti-p-Akt2 (Ser474, CST, #8599), anti-t-Akt3 (Biorbyt, #orb6787), anti-p-Akt3 (Ser472, Biorbyt, #orb6790), anti-t-mTOR (CST, #2983), anti-p-mTOR (Ser2448, CST, #5536), anti-t-p70S6K (SAB, #21276) and anti-p-p70S6K (Ser424, SAB, #21276) and anti-β-actin (CST, #4970) as an internal control.

Techniques: Over Expression, Concentration Assay, Western Blot, De-Phosphorylation Assay, Negative Control

Journal:

Article Title: Effect of sensory disuse on geniculate afferents to cat visual cortex

doi:

Figure Lengend Snippet: Computer reconstructions of PHA-L immunostained axonal arbors in area 17 in a normal P40 animal. Row A shows the arbors as originally reconstructed in the coronal plane, and row B shows the arbors from a surface view after a 90-deg rotation along the dorsoventral axis. The total length of the terminal arborization in layer IV has been computed distally to the small gap in the main axonal trunk, well visible in arbor NN22 (see Materials and methods). The arrowheads indicate the boundaries of the layer IV. VD and AP: ventrodorsal and anteroposterior direction along the lateral gyrus.

Article Snippet: The anterograde tracer Phaseolus lectin, PHA-L (Vector Labs., Burlingame, CA; 2.5% solution in 0.1 M sodium phosphate buffered saline, pH 8; Gerfen & Sawchenko, 1984 ), was injected into the LGN in kittens between the ages of P25 and P28, in all cases prior to the deprivation in BD animals.

Techniques:

Journal:

Article Title: Effect of sensory disuse on geniculate afferents to cat visual cortex

doi:

Figure Lengend Snippet: Computer reconstructions of PHA-L immunostained geniculocortical arbors reconstructed in animals binocularly deprived for 1 week. Arbors are shown in coronal view (A), and in surface view (B). Abbreviations and symbols are as in Fig. 1.

Article Snippet: The anterograde tracer Phaseolus lectin, PHA-L (Vector Labs., Burlingame, CA; 2.5% solution in 0.1 M sodium phosphate buffered saline, pH 8; Gerfen & Sawchenko, 1984 ), was injected into the LGN in kittens between the ages of P25 and P28, in all cases prior to the deprivation in BD animals.

Techniques:

Journal:

Article Title: Effect of sensory disuse on geniculate afferents to cat visual cortex

doi:

Figure Lengend Snippet: Computer reconstructions of PHA-L immunostained geniculocortical arbors reconstructed in animals binocularly deprived for 2 weeks. Arbors are shown in coronal view (A), and in surface view (B). Abbreviations and symbols are as in Fig. 1.

Article Snippet: The anterograde tracer Phaseolus lectin, PHA-L (Vector Labs., Burlingame, CA; 2.5% solution in 0.1 M sodium phosphate buffered saline, pH 8; Gerfen & Sawchenko, 1984 ), was injected into the LGN in kittens between the ages of P25 and P28, in all cases prior to the deprivation in BD animals.

Techniques: