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. 2021 Nov 17;11:756148. doi: 10.3389/fonc.2021.756148

Figure 2.

Figure 2

Lnc-THOR shRNA or KO induces NSCLC cell apoptosis. Lnc-THOR shRNA (“sh-S1” and “sh-S2”, two different sequences)-expressing pCan-1 cells, the CRISPR/Cas9-edited Lnc-THOR KO pCan-1 cells (“koTHOR”), or pCan-1 cells with the scramble control shRNA plus the Cas9-KO empty vector (“shC+koC”), were established and cultured for applied time periods; The relative caspase-3 activity (A), expression of the apoptosis-associated proteins (B, C) and ssDNA contents [ELISA assays, (D)] were tested; Mitochondrial depolarization was tested by JC-1 green monomer accumulation (E); Cell apoptosis was tested by nuclear TUNEL staining assay (F). Primary NSCLC cells (pCan-2 and pCan-3, derived from two different patients) or established cell lines (A549 and H1299), stably expressing the “sh-S1” Lnc-THOR shRNA or the scramble control shRNA (“shC”), were established and cultured for applied time periods; The relative caspase-3 activity (G), JC-1 green monomer intensity (H) and the TUNEL-positive nuclei ratio (I) were tested similarly. “Pare” stands for the parental control cells. Data were presented as mean ± standard deviation (SD, n=5). *P < 0.05 vs. “Pare”/”shC” cells. The experiments were repeated five times, with similar results obtained. Scale Bar = 100 μm (D, E).