sj-docx-1-cre-10.1177_02692155241235336 - Supplemental material for Transcranial direct current stimulation for upper extremity motor dysfunction in poststroke patients: A systematic review and meta-analysis

Supplemental material, sj-docx-1-cre-10.1177_02692155241235336 for Transcranial direct current stimulation for upper extremity motor dysfunction in poststroke patients: A systematic review and meta-analysis by Xian Tang, Nan Zhang, Zhiyuan Shen, Xin Guo, Jun Xing, Shujuan Tian and Yuan Xing in Clinical Rehabilitation</p

Human Elp3 participates in regulation the transcription of <i>SSA3</i> in yeast and its HAT domain is essential for this function.

<p>The plasmids containing <i>hELP3</i>, <i>hELP3/HAT</i>⁻ were transformed into the <i>elp3Δ</i> strain, transformant cells were heat-shocked for 10 min at 37°C (HS), or left at 30°C (NHS), and chromatin immunoprecipitation assays with anti-hElp3 antibody were carried out. Immunoprecipitated DNA was analyzed by polymerase chain reaction using primers specific to the open reading frame (ORF) and 3′ untranslated region (UTR) of the <i>SSA3</i> gene. Products were analyzed by agarose gel electrophoresis followed by ethidium bromide staining. Input: total input DNA or sonicated genome DNA as template, The “input” sample is an aliquot of chromatin that has not been immunoprecipitated but has been crosslinked, sonicated, and processed in a manner identical to the experimental samples; a-hElp3: hElp3 antibody immunoprecipitated DNA as template; NoAb: primary antibody was omitted.</p

Probe and primer sequence used in this paper.

<p>Probe and primer sequence used in this paper.</p

Expression of <i>ELP3</i> gene in HeLa cells.

<p>A: Northern blot products of exogenous gene. The arrows indicate the bands of Northern blot products of exogenous gene (upper arrow), <i>ELP3</i> (middle arrow) and the internal control of <i>GAPDH</i> (lower arrow). B: Relative expression level of Elp3 normalized to <i>GAPDH</i> via photodensitometric analysis of Northern blot products. A significant difference is indicated by * (p<0.01). Non-transfected HeLa cells were used as control.</p

Human Elp3 can complement the growth defect and slow activation of <i>SSA3</i> in <i>elp3Δ</i> cells and the HAT domain is essential for this function.

<p>A: Complementtation tests of <i>hELP3</i>, <i>hELP3/HAT⁻</i> in the <i>elp3Δ</i> strain. The plasmids <i>hELP3</i>, <i>hELP3/HAT⁻</i> were transformed into the <i>elp3Δ</i> strain, and the transformants were plated in a dilution series onto SD-Ura medium and grown at 30°C for 2 days, the transformants were checked for temperature sensitivity after incubation at 39°C for 4 days. B: Complementation of the slow activation of <i>SSA3</i> in <i>elp3Δ</i> cells. Cells were grown at 26°C before transfer to 37°C to activate the <i>SSA3</i> gene. Total RNA was extracted for RT-PCR at 0 or 10 min after heat shocked. The RT-PCR products derived from transcripts of <i>SSA3</i> and <i>ACT1</i> (the internal control) are shown in the left panel. The panels on the right show the results of the quantitative densitometric analyses of the band intensities at each time-point.</p

Hsp70 protein level in HeLa cells.

<p>A: Western blot results of Hsp70 protein level in HeLa cells. HeLa cells were heat shocked for 1 Hour at 42°C and recovered at 37°C for 8–16 hours. Total cell extracts (1×10⁶) were subjected to Western-blot analyses for Hsp70 level. B: Quantitative densitometric analyses of band intensities. A significant difference is indicated by either *(p<0.01) or # (p<0.05). Non-transfected HeLa cells were used as control.</p